185 entries found for : control-conjugates
Madrid LI, Bandhavkar S, Hafey K, Jimenez-Martin J, Milne M, Coulson EJ, Jhaveri DJ (2022) Stimulation of the muscarinic receptor M4 activates quiescent neural precursor cells and ameliorates medial septum cholinergic lesion-induced impairments in adult hippocampal neurogenesis. bioRxiv 2022.08.25.505357. doi: 10.1101/2022.08.25.505357
Objective: To investigate the contribution of basal forebrain medial septum (MS) and diagonal band of Broca (DBB) cholinergic neurons that innervate the hippocampus and the identity of the cholinergic receptor(s) that regulate the production and maturation of new neurons.
Summary: This work reveals stage-specific roles of cholinergic signaling in regulating functionally relevant adult hippocampal neurogenesis.
Usage: Medial septum cholinergic lesion was achieved by infusion of mu p75-SAP (0.4 µg/µl). Rabbit IgG-SAP (0.4 µg/µl) was used as control.
Krieger JP, Asker M, Van der Velden P, Börchers S, Richard JE, Maric I, Longo F, Singh A, De Larigue G, Skibicka KP (2022) Neural pathway for gut feelings: vagal interoceptive feedback from the gastrointestinal tract is a critical modulator of anxiety-like behavior. Biological Psychiatry in press. doi: 10.1016/j.biopsych.2022.04.020
Objective: To determine how the sensing of gastrointestinal state affects anxiety.
Summary: Vagal sensory signals from the gastrointestinal tract are critical for baseline and feeding-induced tuning of anxiety via the central amygdala in rats. The article results suggest vagal gut-brain signaling as a target to normalize interoception in anxiety.
Usage: 1.5 ul of CCK-SAP or Blank-SAP were delivered into each nodose ganglion at 250 ng/ul.
Nguyen KL, Lamerand SR, Deshpande RP, Taylor BK (2021) Featured Article: Selective ablation of IB4+ primary afferent neurons reduces mechanical and cold hyperalgesia in an EAE mouse model of multiple sclerosis. Targeting Trends 22
Jimenez-Martin J, Potapov D, Potapov K, Knöpfel T, Empson RM (2021) Cholinergic modulation of sensory processing in awake mouse cortex. Sci Rep 11(1):17525. doi: 10.1038/s41598-021-96696-8
Objective: To decipher the timing and significance of acetylcholine actions.
Summary: Study provides new insights into how the cortex processes sensory information and how loss of acetylcholine, for example in Alzheimer’s Disease, disrupts sensory behaviours.
Usage: Focal cortical injection of mu p75-SAP or Rabbit IgG-SAP (1.7 mg/ml, 0.3 µl total volume, rate 0.075 µl/minute).Related Products: mu p75-SAP (Cat. #IT-16), Rabbit IgG-SAP (Cat. #IT-35)
Maejima Y, Yokota S, Shimizu M, Horita S, Kobayashi D, Hazama A, Shimomura K (2021) The deletion of glucagon-like peptide-1 receptors expressing neurons in the dorsomedial hypothalamic nucleus disrupts the diurnal feeding pattern and induces hyperphagia and obesity. Nutr Metab (Lond) 18(1):58. doi: 10.1186/s12986-021-00582-z
Summary: Feeding rhythm disruption contributes to the development of obesity. GLP-1 receptors (GLP-1R) are expressed in the dorsomedial hypothalamic nucleus (DMH) which are known to be associated with thermogenesis and circadian rhythm development. These findings suggest that GLP-1R expressing neurons in the DMH may mediate feeding termination.
Usage: Exenatide-SAP targets GLP-1R expressing cells. Injections of 0.1 μg/0.5 μl Ex4-SAP or 0.1 μg/0.5 μl Blank-SAP (control) were administered into the DMH.
Loftén A, Adermark L, Ericson M, Söderpalm B (2021) An acetylcholine-dopamine interaction in the nucleus accumbens and its involvement in ethanol's dopamine-releasing effect. Addict Biol 26(3):e12959. doi: 10.1111/adb.12959
Summary: Basal extracellular levels of dopamine within the nucleus accumbens are not sustained by muscarinic acetylcholine, whereas accumbal Cholinergic interneurons-ACh are involved in mediating ethanol-induced dopamine release.
Usage: Anti-ChAT-SAP or Rabbit IgG-SAP were infused at a flow rate of 0.05 μl/min for 10 min giving a total volume of 0.5 μl.Related Products: Anti-ChAT-SAP (Cat. #IT-42), Rabbit IgG-SAP (Cat. #IT-35)
Wang Z, Jiang C, Yao H, Chen O, Rahman S, Gu Y, Zhao J, Huh Y, Ji RR (2021) Central opioid receptors mediate morphine-induced itch and chronic itch via disinhibition. Brain 144(2):665-681. doi: 10.1093/brain/awaa430
Summary: Itch is a common side effect of opioids, particularly as a result of epidural or intrathecal administration. Notably, morphine-elicited itch was suppressed by intrathecal administration of NPY and abolished by spinal ablation of GRPR+ neurons with intrathecal injection of Bombesin-SAP.
Usage: For ablation of GRPR+ neurons, mice were given an intrathecal injection of 400 ng Bombesin-SAP or Blank-SAP (control) 10 days before behavioral testing.Related Products: Bombesin-SAP (Cat. #IT-40), Blank-SAP (Cat. #IT-21)
Marquez J, Dong J, Dong C, Tian C, Serrero G (2021) Identification of prostaglandin F2 receptor negative regulator (PTGFRN) as an internalizable target in cancer cells for antibody-drug conjugate development. PLoS One 16(1):e0246197. doi: 10.1371/journal.pone.0246197
Summary: PTGFRN is a cell-surface protein that is upregulated in certain cancer types, including head and neck and, notably, pediatric medulloblastoma, an aggressive cancer with limited therapeutic options. With the selection of the mouse monoclonal antibody 33B7, the authors identified PTGFRN as a potential therapy target, and show that it is internalized by incubation with 33B7. Purified 33B7 antibody was sent to Advanced Targeting Systems where saporin was directly conjugated to the Fc region of 33B7 using their proprietary cleavable linker.
Usage: In a 96-well plate, 2000 cells/well were plated in triplicate in 100 μL of DMEM/F12 medium supplemented with 2.5% FBS, 0.4 ug/ml 33B7 antibody, and 0.9ug/ml of Fab-ZAP mouse. As an isotype control, cells were incubated with mouse Fab IgG-SAP as control (instead of 33B7) and Fab-ZAP.Related Products: Fab-ZAP mouse (Cat. #IT-48), Fab IgG-SAP (Cat. #IT-67), Custom Conjugates
Oti T, Satoh K, Uta D, Nagafuchi J, Tateishi S, Ueda R, Takanami K, Young LJ, Galione A, Morris JF, Sakamoto T, Sakamoto H (2021) Oxytocin influences male sexual activity via non-synaptic axonal release in the spinal cord. Curr Biol 31(1):103-114.e5. doi: 10.1016/j.cub.2020.09.089
Summary: Oxytocin directly activates SEG (Spinal Ejaculation Generator)/GRP (Gastrin-Releasing Peptide) neurons via OXTRs (Oxytocin Receptors) and influences male sexual function in the rat lumbar spinal cord.
Usage: Oxytocin-SAP (4 or 40 ng) was infused slowly into the L3 and L4 spinal cord. Blank-SAP was used as control.Related Products: Oxytocin-SAP (Cat. #IT-46), Blank-SAP (Cat. #IT-21)
Tapa S, Wang L, Francis Stuart SD, Wang Z, Jiang Y, Habecker BA, Ripplinger CM (2020) Adrenergic supersensitivity and impaired neural control of cardiac electrophysiology following regional cardiac sympathetic nerve loss. Sci Rep 10:18801. doi: 10.1038/s41598-020-75903-y
Summary: The authors present a novel mouse model of regional cardiac sympathetic hypo-innervation utilizing Anti-DBH-SAP.
Usage: Either 5μL of 40 ng/μL Anti-DBH-SAP or Mouse IgG-SAP (control) was applied three times directly to the exposed apical/anterior surface of the heart.Related Products: Anti-DBH-SAP (Cat. #IT-03), Mouse IgG-SAP (Cat. #IT-18)
Liu X, Wang D, Wen Y, Zeng L, Li Y, Tao T, Zhao Z, Tao A (2020) Spinal GRPR and NPRA contribute to chronic itch in a murine model of allergic contact dermatitis. J Invest Dermatol 140(9):1856-1866.e7. doi: 10.1016/j.jid.2020.01.016
Objective: The authors investigated the peripheral and spinal mechanisms responsible for prolonged itch in a mouse model of allergic contact dermatitis (ACD) induced by squaric acid dibutylester (SADBE).
Summary: Targeting gastrin-releasing peptide receptor (GRPR) and natriuretic peptide receptor A (NPRA) may provide effective treatments for ACD associated chronic pruritus.
Usage: A single dose of Bombesin-SAP (400 ng) and Blank-SAP (400 ng) or two doses of Nppb-SAP (BNP-SAP; 650 ng) and Blank-SAP (650 ng) were administered via intrathecal injection.Related Products: Bombesin-SAP (Cat. #IT-40), Nppb-SAP (Cat. #IT-69), Blank-SAP (Cat. #IT-21)
Kiguchi N, Uta D, Ding H, Uchida H, Saika F, Matsuzaki S, Fukazawa Y, Abe M, Sakimura K, Ko MC, Kishioka S (2020) GRP receptor and AMPA receptor cooperatively regulate itch-responsive neurons in the spinal dorsal horn. Neuropharmacology 170:108025. doi: 10.1016/j.neuropharm.2020.108025
Objective: To investigate the mechanisms for the activation of itch-responsive GRPR+ neurons in the spinal dorsal horn (SDH).
Summary: These findings demonstrate that GRP and glutamate cooperatively regulate GRPR+ AMPAR+ neurons in SDH, mediating itch sensation. GRP–GRPR and the glutamate–AMPAR system may play pivotal roles in the spinal transmission of itch in rodents and nonhuman primates.
Usage: Bombesin-SAP and Blank-SAP were administered i.t. (5 μg/5 μl).Related Products: Bombesin-SAP (Cat. #IT-40), Blank-SAP (Cat. #IT-21)
Harris RBS (2020) Loss of leptin receptor-expressing cells in the hindbrain decreases forebrain leptin sensitivity. Am J Physiol Endocrinol Metab 318(5):E806-E816. doi: 10.1152/ajpendo.00020.2020
Objective: This study tested whether loss of hindbrain leptin receptor signaling changed sensitivity to forebrain leptin.
Summary: Loss of VMH (ventromedial nucleus of hippocampus) leptin receptor-expressing cells prevents weight loss. The integrated response between the hindbrain and forebrain is heavily dependent upon leptin activity in the VMH.
Usage: To test forebrain leptin sensitivity Leptin-SAP and Blank-SAP rats received third ventricle injections of 0, 0.05, 0.1, 0.25 or 0.5 μg leptin.Related Products: Leptin-SAP (Cat. #IT-47), Blank-SAP (Cat. #IT-21)
Qian L, Kasas L, Milne MR, Rawashdeh O, Marks N, Sharma A, Bellingham MC, Coulson EJ (2020) Cholinergic basal forebrain degeneration due to obstructive sleep apnoea increases Alzheimer’s pathology in mice. bioRxiv 2020.03.12.989848. doi: 10.1101/2020.03.12.989848
Usage: bilateral injections of urotensin II-saporin (UII-SAP; 0.07 μg/μL per site – unless stated otherwise; generous gift from Advanced Targeting Systems)Related Products: Custom Conjugates, Blank-SAP (Cat. #IT-21)
Bernabe CS, Caliman IF, Truitt WA, Molosh AI, Lowry CA, Hay-Schmidt A, Shekhar A, Johnson PL (2020) Using loss- and gain-of-function approaches to target amygdala-projecting serotonergic neurons in the dorsal raphe nucleus that enhance anxiety-related and conditioned fear behaviors. J Psychopharmacol 34(4):400-411. doi: 10.1177/0269881119900981
Objective: To investigate the role of amygdala-projecting 5-HT neurons in the DR in innate anxiety and conditioned fear behaviors.
Summary: The studies support the hypothesis that amygdala-projecting 5-HT neurons in the DR represent an anxiety and fear-on network.
Usage: Each rat received two bilateral microinjections per site (100 nL each, 1 μM in artificial cerebrospinal fluid) of either Anti-SERT-SAP or the control Mouse IgG-SAP.Related Products: Anti-SERT-SAP (Cat. #IT-23), Mouse IgG-SAP (Cat. #IT-18)
Zhang F, Huan L, Xu T, Li G, Zheng B, Zhao H, Guo Y, Shi J, Sun J, Chen A (2020) Inflammatory macrophages facilitate mechanical stress-induced osteogenesis. Aging (Albany NY) 12(4):3617-3625. doi: 10.18632/aging.102833
Summary: In a mouse model of distraction osteogenesis (DO), there was significant increase in macrophages in the regeneration area. This suggests that targeting inflammatory macrophages may help to improve clinical bone repair.
Usage: For saporin-mediated depletion of macrophages, DO-surgery-treated mice received an intraventricular (iv) injection of either Mac-1-SAP or Rat IgG-SAP (20µg) once every 3 days.Related Products: Mac-1-SAP mouse/human (Cat. #IT-06), Rat IgG-SAP (Cat. #IT-17)
Goodman RL, He W, Lopez JA, Bedenbaugh MN, McCosh RB, Bowdridge EC, Coolen LM, Lehman MN, Hileman SM (2019) Evidence that the LH surge in ewes involves both neurokinin B–dependent and –independent actions of kisspeptin. Endocrinology 160(12):2990-3000. doi: 10.1210/en.2019-00597
Objective: To determine if NKB is involved in the RCh of the ewe in the LH surge.
Summary: NKB signaling in the RCh increases kisspeptin levels critical for the full amplitude of the LH surge in the ewe, but kisspeptin release occurs independently of retrochiamatic area (RCh) input at the onset of the surge to initiate GnRH secretion.
Usage: Bilaterial injections in the RCh of either NK3-SAP or Blank-SAP.Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Tominaga M, Kusube F, Honda K, Komiya E, Takahashi N, Naito H, Suga Y, Takamori K (2019) OP11: Role of spinal cholecystokinin receptor 2 in alloknesis models. Itch 4:1-62. doi: 10.1097/itx.0000000000000030
Objective: To determine the detailed molecular and cellular mechanisms that induce alloknesis via the spinal CCK2 receptor.
Summary: Ablation of spinal CCK receptor-expressing cells by i.t. injection of CCK-SAP attenuated CCK8S-induced alloknesis in comparison with Blank-SAP control mice.
Usage: Intrathecal injectionRelated Products: CCK-SAP (Cat. #IT-31), Blank-SAP (Cat. #IT-21)
Seamon M, Ahn W, Li AJ, Ritter S, Harris RBS (2019) Leptin receptor-expressing neurons in ventromedial nucleus of the hypothalamus contribute to weight loss caused by fourth ventricle leptin infusions. Am J Physiol Endocrinol Metab 317(4):E586-E596. doi: 10.1152/ajpendo.00205.2019
Objective: To test the importance of VMH leptin responsiveness in mediating weight loss caused by fourth ventricle leptin infusion.
Summary: Leptin did not inhibit food intake and respiratory exchange ratio in rats treated with Leptin-SAP. VMH leptin receptors do not play a significant role in maintaining energy balance in basal conditions, but limit weight gain during positive energy balance.
Usage: Bilateral VMH 75-nl injections of 260 ng/ml of Leptin-SAP or Blank-SAP.Related Products: Leptin-SAP (Cat. #IT-47), Blank-SAP (Cat. #IT-21)
Pan H, Fatima M, Li A, Lee H, Cai W, Horwitz L, Hor CC, Zaher N, Cin M, Slade H, Huang T, Xu XZS, Duan B (2019) Identification of a spinal circuit for mechanical and persistent spontaneous itch. Neuron 103(6):1135-1149.e6. doi: 10.1016/j.neuron.2019.06.016
Objective: To identify a spinal circuit for mechanical and persistent spontaneous itch.
Summary: Findings indicate excitatory interneurons expressing Urocortin 3::Cre (Ucn3+) in the dorsal spinal cord as a valid cellular target for future therapeutic interventions against chronic itch, without affecting normal touch.
Usage: To ablate spinal GRPR+ neurons, mice were given a single intrathecal injection of either Bombesin-SAP or Blank-SAP (400 ng in 10 mL sterile saline). To ablate spinal Npra+ neurons, mice were given a single intrathecal injection of either Nppb-SAP or Blank-SAP (5 mg in 10 mL sterile saline).
Katsurada K, Nakata M, Saito T, Zhang B, Maejima Y, Nandi SS, Sharma NM, Patel KP, Kario K, Yada T (2019) Central glucagon-like peptide-1 receptor signaling via brainstem catecholamine neurons counteracts hypertension in spontaneously hypertensive rats. Sci Rep 9(1):12986. doi: 10.1038/s41598-019-49364-x
Objective: To determine mechanisms for antihypertensive effect of GLP-1R agonists.
Summary: The central GLP-1R signaling via NTS DBH neurons counteracts the development of hypertension in SHR, accompanied by attenuated sympathetic nerve activity.
Usage: Anti-DBH-SAP or Blank-SAP was injected into NTS bilaterally (6 ng/200 nl).Related Products: Anti-DBH-SAP (Cat. #IT-03), Blank-SAP (Cat. #IT-21)
Maejima Y, Yokota S, Horita S, Shimomura K (2019) The undeveloped properties of GABA neurons in the ventral tegmental area promote energy intake for growth in juvenile rats. Sci Rep 9(1):11848. doi: 10.1038/s41598-019-48336-5
Objective: To determine the underlying mechanisms that induce high energy intake (EI) per body weight (BW).
Summary: Undeveloped properties of VTA GABA neurons in juvenile rats can promote higher EI regardless of high or less palatable feeding, and contribute to growth promotion.
Usage: GAT1-SAP or control, Rabbit IgG-SAP, was bilaterally injected (0.025 μg/0.5 μl) into the VTA in eight-week-old adult rats.Related Products: GAT1-SAP (Cat. #IT-32), Rabbit IgG-SAP (Cat. #IT-35)
Sakata D, Uruno T, Matsubara K, Andoh T, Yamamura K, Magoshi Y, Kunimura K, Kamikaseda Y, Furue M, Fukui Y (2019) Selective role of neurokinin B in IL-31–induced itch response in mice. J Allergy Clin Immunol 144(4):1130-1133. doi: 10.1016/j.jaci.2019.06.031
Objective: To examine the physiological significance of neurokinin B in IL-31–induced itch sensation.
Summary: IL-31–induced scratching was unaffected by intrathecal injection of Nppb-SAP. In contrast,treatment with Bombesin-SAP reduced IL-31–induced scratching. Neurokinin B acts upstream of GRP to transmit IL-31–induced itch sensation.
Usage: Intrathecal injection
Acton D, Ren X, DiCostanzo S, Dalet A, Bourane S, Bertocchi I, Eva C, Goulding M (2019) Spinal neuropeptide Y1 receptor-expressing neurons form an essential excitatory pathway for mechanical itch. Cell Reports 28(3):625-639.e6 . doi: 10.1016/j.celrep.2019.06.033
Objective: To determine the central pathway for mechanical itch.
Summary: NPY-Y1 signaling regulates the transmission of innocuous tactile information by establishing biologically relevant thresholds for touch discrimination and mechanical itch reflexes. Neither the evoked nor spontaneous scratching seen following activation of Y1Cre neurons was affected by ablation of the GRPR+ neurons. NK1R+ neuron ablation failed to modulate mechanically evoked itch.
Usage: P28 mice were given a single intrathecal (i.t.) injection of either Bombesin-SAP (400 ng in 5 mL 0.9% sterile saline) to ablate GRPR+ cells or SSP-SAP to ablate NK1r+ neurons (100 ng in 5 mL 0.9% sterile saline). Littermate controls received Blank-SAP (equal mass in 5 mL 0.9% sterile saline).Related Products: Bombesin-SAP (Cat. #IT-40), SSP-SAP (Cat. #IT-11), Blank-SAP (Cat. #IT-21)
Weiner GA, Shah SH, Angelopoulos CM, Bartakova AB, Pulido RS, Murphy A, Nudleman E, Daneman R, Goldberg JL (2019) Cholinergic neural activity directs retinal layer-specific angiogenesis and blood retinal barrier formation. Nat Commun 10(1):2477. doi: 10.1038/s41467-019-10219-8
Objective: To determine which neurons are responsible for angiogenesis and blood retinal barrier formation.
Summary: Anti-ChAT-SAP reduces SAC (starburst amacrine cell) number and inhibits deep-layer angiogenesis.
Usage: Anti-ChAT-SAP or control Rabbit-IgG-SAP were injected intravitreally at P3 and P11 (0.12 mg/mL in PBS).Related Products: Anti-ChAT-SAP (Cat. #IT-42), Rabbit IgG-SAP (Cat. #IT-35)
Nelson TS, Fu W, Donahue RR, Corder GF, Hökfelt T, Wiley RG, Taylor BK (2019) Facilitation of neuropathic pain by the NPY Y1 receptor-expressing subpopulation of excitatory interneurons in the dorsal horn. Sci Rep 9(1):7248. doi: 10.1038/s41598-019-43493-z
Objective: To test the relevance of the NPYY1 spinal population to the development and/or maintenance of acute and neuropathic pain.
Summary: This neuroanatomical and behavioral characterization of Y1R-expressing excitatory interneurons provides compelling evidence for the development of spinally-directed Y1R agonists to reduce chronic neuropathic pain.
Usage: Selectively ablated Y1R-expressing interneurons while sparing the central terminals of primary afferents. Rats received intrathecal injections of either NPY-SAP or control Blank-SAP (1000 ng each).Related Products: NPY-SAP (Cat. #IT-28), Blank-SAP (Cat. #IT-21)
Fatima M, Ren X, Pan H, Slade HFE, Asmar AJ, Xiong CM, Shi A, Xiong AE, Wang L, Duan B (2019) Spinal somatostatin-positive interneurons transmit chemical itch. Pain 160(5):1166-1174. doi: 10.1097/j.pain.0000000000001499
Objective: To further study the cellular identity of spinal interneurons that contribute to itch processing.
Summary: Findings reveal a novel spinal mechanism for sensory encoding of itch perception.
Usage: Npra receptor–expressing spinal cord interneurons were ablated through intrathecal injection of Nppb-SAP (5 μg/10 μL) or control Blank-SAP in lumbar segment 3 to 4. Behavioral analyses were performed 1 week after the toxin injection.Related Products: Nppb-SAP (Cat. #IT-69), Blank-SAP (Cat. #IT-21)
Krajewski-Hall SJ, Miranda Dos Santos F, McMullen NT, Blackmore EM, Rance NE (2019) Glutamatergic neurokinin 3 receptor neurons in the median preoptic nucleus modulate heat-defense pathways in female mice. Endocrinology 160(4):803-816. doi: 10.1210/en.2018-00934
Objective: To characterize the thermoregulatory role of MnPO NK3R neurons in female mice.
Summary: Study suggests that KNDy neurons modulate thermosensory pathways for heat defense indirectly via a subpopulation of glutamatergic MnPO neurons that express NK3R.
Usage: Mice were bilaterally injected with 10 ng NK3-SAP in 100 nL PBS (n = 14) or blank-SAP (n = 8) in the preoptic area adjacent to the MnPO.Related Products: NKB-SAP (Cat. #IT-63), Blank-SAP (Cat. #IT-21)
Eidson LN, Murphy AZ (2019) Inflammatory mediators of opioid tolerance: Implications for dependency and addiction. Peptides 115:51-58. doi: 10.1016/j.peptides.2019.01.003
Objective: To determine what mediates opioid tolerance and alterations in glutamate homeostasis.
Summary: Site-specific lesions of PAG MOR-containing neurons using Dermorphin-SAP significantly reduce the antinociceptive effects of systemic morphine suggesting that PAG MOR is critical for morphine action.
Usage: Rats were injected with 3 pmol of Dermorphin-SAP (Cat. #IT-12) into the PAG. Blank-SAP (Cat. #IT-21) was used as a control.
See: Loyd DR et al. Sex differences in micro-opioid receptor expression in the rat midbrain periaqueductal gray are essential for eliciting sex differences in morphine analgesia. J Neurosci 28:14007-14017, 2008.Related Products: Dermorphin-SAP / MOR-SAP (Cat. #IT-12), Blank-SAP (Cat. #IT-21)
Gao ZR, Chen WZ, Liu MZ, Chen XJ, Wan L, Zhang XY, Yuan L, Lin JK, Wang M, Zhou L, Xu XH, Sun YG (2019) Tac1-expressing neurons in the periaqueductal gray facilitate the itch-scratching cycle via descending regulation. Neuron 101(1):45-59.e9. doi: 10.1016/j.neuron.2018.11.010
Objective: To determine the neural mechanism promoting the itch-scratching cycle.
Summary: Ablation of Tac1+ but not SST+ neurons decreases itch-induced scratching behavior. l/vlPAG Tac1+ neurons Induce Scratching Behavior via a Descending Pathway.
Usage: To ablate the spinal GRPR+ neurons, mice were intrathecally injected with Bombesin-SAP or Control Blank-SAP (400 ng/5 mL).Related Products: Bombesin-SAP (Cat. #IT-40), Blank-SAP (Cat. #IT-21)
Goodman RL, Lopez JA, Bedenbaugh MN, Connors JM, Hardy SL< Hileman SM, Coolen LM, Lehman MN (2018) Evidence that the LH surge in ewes involves both neurokinin B-dependent and -independent actions of kisspeptin. Neuroscience 2018 Abstracts 773.20 / YY14. Society for Neuroscience, San Diego, CA.
Summary: It is generally recognized that kisspeptin plays a key role in induction of the LH surge in sheep and we have reported evidence that neurokinin B (NKB) does so as well. Specifically, disrupting NKB signaling in the retrochiasmatic area (RCh) using either an antagonist to its receptor, NK3R, or lesions of NK3R-containing neurons in the RCh with a saporin conjugate (NK3-SAP) reduced the amplitude of the estrogen-induced LH surge by 50%. Because a KISS1R antagonist (p271) also produced a 50% decrease in surge amplitude, we hypothesized that these two systems are organized in series with NKB actions in the RCh stimulating kisspeptin release. If this is the case, then the combination of NK3R lesions and a KISS1R antagonist should produce the same inhibition as either treatment alone. This experiment tested this prediction using a 2 x 2 design. Breeding season ewes were ovariectomized and immediately given an estradiol (E) implant sc and two progesterone implants (CIDRs) intravaginally that produced luteal phase levels of these steroids. Ewes then received bilateral injections of either NK3-SAP (n=6) or Blank-SAP (n=5) into the RCh. Three weeks later, an artificial follicular phase was produced by inserting four 3 cm long E implants 24 hrs after CIDR removal and either saline or p271 was infused into the lateral ventricle for 16-24 hrs after E implantation; LH was monitored every 2-4 hrs for two days. CIDRs were then reinserted and the protocol repeated in a cross-over design so that all ewes received saline and p271 treatment. In Blank-SAP ewes, p271 decreased the peak of the LH surge from 61.2 ± 7.6 to 27.4 ± 4.6 ng/mL and delayed it 8 hrs (from 26.5 ± 0.5 to 34.1 ± 1.2 hrs post E implantation). The NK3-SAP injections alone decreased the peak of the LH surge to 29.7 ± 10.7 ng/mL compared to Blank-SAP, but the peak was not further inhibited by p271 in these NK3-SAP-treated ewes (24.4 ± 1.4 ng/mL). However, p271 delayed the peak of the LH surge (from 28.8 ± 1.2 to 34.8 ± 2.1 hrs post E implantation) in the ewes injected with NK3-SAP. Based on these results, we propose that kisspeptin has two roles in the LH surge in ewes: it initiates the surge independent of NKB signaling in the RCh, and maintains LH secretion during the surge by a NKB-dependent system.
Qian L, Milne MR, Shepheard S, Rogers ML, Medeiros R, Coulson EJ (2019) Removal of p75 neurotrophin receptor expression from cholinergic basal forebrain neurons reduces amyloid-β plaque deposition and cognitive impairment in aged APP/PS1 mice. Mol Neurobiol 56(7):4639-4652. doi: 10.1007/s12035-018-1404-2
Objective: To investigate the contribution of CBF neuronal p75NTR to the progression of Alzheimer's Disease
Summary: Data indicate that a direct interaction between CBF-expressed p75NTR and Aβ does not contribute significantly to the regulation of Aβ load.
Usage: To lesion CBF neurons, a single infusion of mu p75-SAP or control Rabbit IgG-SAP (0.4 mg/ml) was stereotaxically-injected into the basal forebrain.Related Products: mu p75-SAP (Cat. #IT-16), Rabbit IgG-SAP (Cat. #IT-35)
Acharjee S, Verbeek M, Gomez CD, Bisht K, Lee B, Benoit L, Sharkey KA, Benediktsson A, Tremblay M-E, Pittman QJ (2018) Reduced microglial activity and enhanced glutamate transmission in the basolateral amygdala in early CNS autoimmunity. J Neurosci 38:9019-9033. doi: 10.1523/JNEUROSCI.0398-18.2018
Objective: To identify CNS changes associated with behaviors in multiple sclerosis (MS) patients.
Summary: The data from this study reveal increased synaptic activity and spine density in early stages of experimental autoimmune encephalomyelitis (an animal model of MS) in the basolateral amygdala.
Usage: Mac-1-SAP mouse/human or Rat-IgG-SAP (control) was injected unilaterally in the BLA (1 ug/1 ul).Related Products: Mac-1-SAP mouse/human (Cat. #IT-06), Rat IgG-SAP (Cat. #IT-17)
Oliveira LM, Falquetto B, Moreira TS, Takakura AC (2018) Orexinergic neurons are involved in the chemosensory control of breathing during the dark phase in a Parkinson's disease model. Exp Neurol 309:107-118. doi: 10.1016/j.expneurol.2018.08.004
Objective: To determine the involvement of orexin cells from the lateral hypothalamus/perifornical area (LH/PeF) on breathing.
Summary: The degeneration of orexinergic neurons in this model of PD can be related to impaired chemoreceptor function in the dark phase.
Usage: For lesions of LH/PeF, two injections of Orexin-B-SAP or Rabbit IgG-SAP (100 ng/μl) were made into the lateral hypothalamus / perifornical area (LH/PeF).Related Products: Orexin-B-SAP (Cat. #IT-20), Rabbit IgG-SAP (Cat. #IT-35)
Holschbach MA, Vitale EM, Lonstein JS (2018) Serotonin-specific lesions of the dorsal raphe disrupt maternal aggression and caregiving in postpartum rats. Behav Brain Res 348:53-64. doi: 10.1016/j.bbr.2018.04.008
Objective: To determine the effects of behavioral modifications associated with early motherhood by permanently disrupting serotonin signaling at one of its primary sources, the dorsal raphe (DR).
Summary: Prepartum serotonin-specific lesions of the DRdm impaired maternal aggression. Larger postpartum DR serotonin lesions affected both aggression and caregiving. DR serotonin lesions did not affect postpartum anxiety.
Usage: 1 μL of 0.1M anti-SERT-SAP or control Mouse IgG-SAP was slowly infused into the DR.Related Products: Anti-SERT-SAP (Cat. #IT-23), Mouse IgG-SAP (Cat. #IT-18)
Zhang Q, De Corte B, Jung D, Kim Y, Geerling J, Narayanan N (2018) Cholinergic modulation targeting medial prefrontal cortex leads to behavior deficit in interval timing task. Neurology 90 (15 Supplement):P5.195.
Objective: To determine the effect of cholinergic lesion targeting medial prefrontal cortex on interval timing behavior.
Summary: Mice receiving medial prefrontal mu-p75-saporin injection performed poorly compared to control mice in interval timing task. Cholinergic lesion targeting medial prefrontal cortex caused interval timing behavior deficit in wild type mice.
Usage: mu-p75-SAP, a toxin targeting cholinergic neurons, into the bilateral medial prefrontal cortical regions of wild type mice pre-trained in interval timing task. Control mice (also pre-trained in interval timing task, n=8) received stereotactic injection of Rabbit IgG-SAP.Related Products: mu p75-SAP (Cat. #IT-16), Rabbit IgG-SAP (Cat. #IT-35)
Strichartz G, Khasabov S, Barr T, Wang J, Simone D (2018) Modulation of chronic post-thoracotomy pain by NK-1 neurons in the rostral ventromedial medulla is not paralleled by changes spinal MAPKinase activation. J Pain 19:S14. doi: 10.1016/j.jpain.2017.12.065
Objective: To evaluate SSP-SAP in treatment of tactile hypersensitivity for Chronic Post-Thoractotomy Pain (CPTP).
Summary: SSP-SAP 3 weeks before thoracotomy and rib retraction (TRR) was able to completely prevent CPTP, assayed by tactile hypersensitivity.
Usage: Ablation of Neurokinin-1 receptor (NK-1R)- expressing neurons in the rat rostral ventromedial medulla (RVM), by micro-injection of the specific neurotoxin SSP-SAP. No effect with treatment of control, Blank-SAP (IT-21).Related Products: SSP-SAP (Cat. #IT-11), Blank-SAP (Cat. #IT-21)
Turnbull MT, Boskovic Z, Coulson EJ (2018) Acute down-regulation of BDNF signaling does not replicate exacerbated amyloid-β levels and cognitive impairment induced by cholinergic basal forebrain lesion. Front Mol Neurosci 11:51. doi: 10.3389/fnmol.2018.00051
Objective: To determine if degeneration of BFCNs causes a decrease in neurotrophin levels in innervated brain areas, which in turn promotes the development of Amyloid beta pathology and cognitive impairment.
Summary: Lesion of septo-hippocampal BFCNs in a pre-symptomatic transgenic amyloid AD mouse model (APP/PS1 mice) increases soluble Ab levels in the hippocampus, and induces cognitive deficits in a spatial memory task that are not seen in either unlesioned APP/PS1 or non-transgenic littermate control mice. Cognitive decline and Amyloid-beta pathology induced by cholinergic basal forebrain neuron loss occur independent of dysfunctional neuronal BDNF signaling, and may therefore be directly underpinned by reduced cholinergic neurotransmission.
Usage: To lesion BFCNs, a single infusion of murine p75-SAP or control rabbit IgG-SAP (0.4 mg/ml) was stereotaxically injected into the basal forebrain.Related Products: mu p75-SAP (Cat. #IT-16), Rabbit IgG-SAP (Cat. #IT-35)
Lee SJ, Jokiaho AJ, Sanchez-Watts G, Watts AG (2018) Catecholaminergic projections into an interconnected forebrain network control the sensitivity of male rats to diet-induced obesity. Am J Physiol Regul Integr Comp Physiol 314(6):R811-R823. doi: 10.1152/ajpregu.00423.2017
Objective: To investigate the role of hindbrain catecholamine neuron pathways and their contribution to long-term energy homeostasis by controlling obesogenic sensitivity to a high-fat, high sucrose choice diet.
Summary: The authors show that catecholamine neurons (primarily in the VLM and NTS) convey essential feedback signals to enable long-term adaptive control of energy metabolism when animals consume a predominantly carbohydrate diet. This is the first report specifically associating this projection system with the long-term control of adiposity.
Usage: Catecholaminergic projections to the PVH and related parts of the forebrain were lesioned with bilateral injections each consisting of 42 ng/200 nL of Anti-DBH-SAP or equimolar amounts of control Mouse IgG-SAP.Related Products: Anti-DBH-SAP (Cat. #IT-03), Mouse IgG-SAP (Cat. #IT-18)
Mu D, Sun Y-G (2017) Itch induces conditioned place aversion in mice. Neuroscience Letters 658:91-96.. doi: 10.1016/j.neulet.2017.08.046
Summary: Consistently, ablation of itch‐specific neurons that express gastrin‐releasing peptide receptor in the spinal cord also abolished itch‐induced Conditioned Place Aversion (CPA), confirming that itch‐induced CPA is dependent on the spinal itch circuit.
Usage: Mice were given a single intrathecal injection (400 ng/5 μl each) of Bombesin-SAP (Cat. #IT-40) or Control Blank-SAP (Cat. #IT-21).
Fu C, Xue J, Wang R, Chen J, Ma L, Liu Y, Wang X, Guo F, Zhang Y, Zhang X, Wang S (2017) Chemosensitive Phox2b-expressing neurons are crucial for hypercapnic ventilatory response in the nucleus tractus solitarius. J Physiol 595:4973-4989.. doi: 10.1113/JP274437
Objective: To investigate whether paired-like homeobox 2b (Phox2b)-expressing NTS neurons are recruited in hypercapnic ventilatory response (HCVR) and whether these neurons exhibit intrinsic chemosensitivity.
Summary: Respiratory deficits caused by injection of SSP-SAP into the NTS are attributable to proportional lesions of CO2/H+-sensitive Phox2b-expressing neurons.
Usage: Two protocols were applied for SSP-SAP injections. In immunostaining experiments, to determine how many Phox2b-containing cells were destroyed, a total volume of 100 nl of PBS containing 6 ng of SSP-SAP (3 ng in 50 nl; 2 injections) was injected into one side of the NTS and the contralateral NTS was used as a control (no injection). For in vivo experiments, to determine whether loss of Phox2b cells led to impaired HCVR, bilateral injections with a total volume of 200 nl of PBS containing 6 ng (1.5 ng in 50 nl per injection; 2 injections per side) or 12 ng (3 ng in 50 nl per injection; two injections per side) of toxin into NTS. Breathing was studied in conscious, freely moving mice treated with SSP-SAP and Blank-SAP.
Havelin J, Imbert I, Sukhtankar D, Remeniuk B, Pelletier I, Gentry J, Okun A, Tiutan T, Porreca F, King T (2017) Mediation of movement-induced breakthrough cancer pain by IB4-binding nociceptors in rats. J Neurosci 37:5111-5122.. doi: 10.1523/JNEUROSCI.1212-16.2017
Objective: To define a novel preclinical measure of movement-induced breakthrough pain (BTP) that is observed in the presence of morphine controlling ongoing pain.
Summary: Novel compounds targeting IB4-binding nociceptors may improve pain management for cancer pain patients and other patient populations suffering from BTP that is inadequately treated by currently available medications.
Usage: To determine the effect of eliminating input from IB4-binding fibers, separate groups of rats received spinal administration of IB4-SAP or the control, Blank-SAP (3.2 mcg/20 mcl saline) followed by a 10 mcl flush of saline. Movement of an air bubble placed between drug solution and saline was used to monitor progress of the injection.
Ye J-H, Fu R, He W (2017) The rostromedial tegmental nucleus and alcohol addiction. Oncotarget 8:18624-18625.. doi: 10.18632/oncotarget.15822
Summary: The authors discuss their work with Dermorphin-SAP (Cat. #IT-12) and their demonstration that damage of RMTg MOR-expressing GABAergic neurons by Dermorphin-SAP increased the intake and preference for alcohol, boosted the expression and slowed down the extinction of alcohol conditioned place preference, and increased locomotion. Microinjection of DS into the RMTg substantially reduced the number of RMTg cells. Importantly, the rats that received DS injection elevated their alcohol intake and preference compared to those that received an injection of Blank-SAP (Cat. #IT-21), which did not cause neuronal damage.
Kelly SC, Nelson PT, Counts SE (2017) Featured Article: The locus coeruleus: a potential link between cerebrovascular and neuronal pathology in Alzheimer’s disease. Targeting Trends 18.
Turnbull M, Coulson E (2017) Cholinergic basal forebrain lesion decreases neurotrophin signaling without affecting tau hyperphosphorylation in genetically susceptible mice. J Alzheimers Dis 55:1141-1154.. doi: 10.3233/JAD-160805
Summary: Alzheimer’s disease(AD) is a progressive, irreversible neurodegenerative disease that destroys memory and cognitive function. Aggregates of hyperphosphorylated tau protein are a prominent feature in the brain of patients with AD, and area major contributor to neuronal toxicity and disease progression. However, the factors that initiate the toxic cascade that results in tau hyperphosphorylation in AD are unknown. The authors investigated whether degeneration of basal forebrain cholinergic neurons (BFCNs) and/or resultant decrease in neurotrophin signaling cause aberrant tau hyperphosphorylation. Two-month-old male and female pR5 mice were infused with murine p75-SAP (Cat. #IT-16) at a concentration of 0.4 mg/ml or 0.4 mg/ml of control Rabbit IgG-SAP (Cat. #IT-35) using a 30G needle attached to a 5 ml Hamilton syringe and pump. The needle was lowered into the medial septum according to coordinates in a mouse brain atlas, and the toxin was infused at a rate of 0.4 ul/min (1.5 u total volume). The results reveal that the loss of BFCNs in pre-symptomatic pR5 tau transgenic mice results in a decrease in hippocampal brain-derived neurotrophic factor levels and reduced TrkB receptor activation. However, there was no exacerbation of the levels of phosphorylated tau or its aggregation in the hippocampus of susceptible mice. Furthermore the animals’ performance in a hippocampal-dependent learning and memory task was unaltered, and no changes in hippocampal synaptic markers were observed. This suggests that tau pathology is likely to be regulated independently of BFCN degeneration and the corresponding decrease in hippocampal neurotrophin levels, although these features may still contribute to disease etiology.
Kaushal R, Taylor B, Jamal A, Zhang L, Ma F, Donahue R, Westlund K (2016) GABA-A receptor activity in the noradrenergic locus coeruleus drives trigeminal neuropathic pain in the rat; contribution of NAα1 receptors in the medial prefrontal cortex. Neuroscience 334:148-159. doi: 10.1016/j.neuroscience.2016.08.005
Summary: The goal of this study was to investigate the role of the locus coeruleus (LC) in a rat orofacial pain model of trigeminal neuropathy induced by chronic constrictive injury of the infraorbital nerve (CCI-ION). Mechanical thresholds to von Frey filaments were tested on whisker pads to evaluate neuropathic pain behavior; pain was indicated by development of mechanical hypersensitivity. Noradrenergic (NA) neurons were lesioned with 5-mcg injections of Anti-DBH-SAP (Cat. #IT-03) into the left lateral ventricle. Mouse-IgG-SAP (Cat. #IT-18) was used as a control. After ablation of NA neurons there was a notable increase in the mechanical threshold compared to control animals. They also targeted coeruleotrigeminal NA neurons by injecting Anti-DBH-SAP into the trigeminal brainstem nuclei bilaterally in one animal and saw similar results. Injecting a GABAA receptor antagonist into the LC after injury had an inhibitory effect on nerve injury induced hypersensitivity. Injection of a NAα1 receptor antagonist, but not a NAα2 receptor antagonist, into the medial prefrontal cortex (mPFC) alleviates mechanical hypersensitivity. They conclude that GABAA-mediated activation of NA neurons during CCI-ION can facilitate hypersensitivity through NAα1 receptors in the mPFC, and that the LC is a chronic pain generator.
Young J (2016) Hunger, thirst, sex, and sleep: How the brain controls our passions. Rowman & Littlefield Publishers.
Kumar J, Rajkumar R, Lee L, Dawe G (2016) Nucleus incertus contributes to an anxiogenic effect of buspirone in rats: Involvement of 5-HT1A receptors. Neuropharmacology 110:1-14. doi: 10.1016/j.neuropharm.2016.07.019
Summary: The nucleus incertus (NI) is involved in stress and anxiety responses. The NI is a cluster of GABAergic neurons in the brainstem, and coexpresses CRF, 5-HT1A and D2 receptors. Buspirone is a partial agonist of 5-HT1A receptors, an antagonist of D2 receptors, and activates the NI. Buspirone is an anti-anxiety drug, but preclinical studies showed that it induces anxiety at high doses. To see if the NI is necessary for the anxiogenic effects of high doses of buspirone, rats were bilaterally injected with 86 ng of CRF-SAP (Cat. #IT-13) into the NI. Blank-SAP (Cat. #IT-21) was used as a control. NI lesioning alone had an anxiogenic effect in several anxiety screening tests compared to sham-lesioned rats, which suggests that the NI reduces anxiety physiologically. Lesioning with CRF-SAP reduced the anxiogenic effects of intra-NI injections of buspirone. No significant difference in the anxiety screening tests resulted from injecting quinpiole, a D2 agonist, which suggests that the 5HT1A receptors in the NI are involved in the anxiogenic effects of buspirone.
Matsuura T, Kawasaki M, Hashimoto H, Yoshimura M, Motojima Y, Saito R, Ueno H, Maruyama T, Ishikura T, Sabanai K, Mori T, Ohnishi H, Onaka T, Sakai A, Ueta Y (2016) Possible involvement of the rat hypothalamo-neurohypophysial/-spinal oxytocinergic pathways in acute nociceptive responses. J Neuroendocrinol 28(6) doi: 10.1111/jne.12396
Summary: It has been suggested that the amplification of GABAergic neurons in the inhibitory system induces the selective inhibition by Oxytocin (OXT) of excitability in the spinal cord, and the pain transmitted from the periphery to the dorsal horn of the spinal cord by this action may be attenuated at the spinal cord level. Rats were injected IT with Oxytocin-SAP (Cat. #IT-46) dissolved in saline (0.06 μg/μl), Blank-SAP (Cat. #IT-21) dissolved in saline (0.06 μg/μl), or saline. Formalin-induced acute nociception activated OXT-containing cells in both the magnocellular and parvocellular divisions of hypothalamus, and that the parvocellular division remains activated longer than the magnocellular division. Acute nociception-induced activation of the hypothalamo-neurohypophysial system caused elevation of plasma OXT levels. In addition, the OXTergic spinal pathway may be involved in pain modulation via OXTRs in the spinal cord.
Matsuura T, Kawasaki M, Hashimoto H, Yoshimura M, Motojima Y, Saito R, Ueno H, Maruyama T, Sabanai K, Mori T, Ohnishi H, Sakai A, Ueta Y (2016) Effects of central administration of oxytocin-saporin cytotoxin on chronic inflammation and feeding/drinking behaviors in adjuvant arthritic rats. Neurosci Lett 621:104-110. doi: 10.1016/j.neulet.2016.04.010
Summary: In the present study, Oxytocin-SAP, which chemically disrupts oxytocin (OXT signaling was administered centrally and an OXT receptor (OXTR) antagonist administered peripherally to determine whether central and peripheral OXT is involved in chronic inflammation and feeding/drinking behavior in adjuvant arthritis (AA) rats. Rats were injected i.t. with Oxytocin-SAP (Cat. #IT-46) or Blank-SAP (Cat. #IT-21) dissolved in saline (0.06 μg/μl). The results demonstrated that the arthritis index values were significantly enhanced and suppression of food intake was transiently attenuated in Oxytocin-SAP treated rats when AA developed, The arthritis index and food intake did not significantly change in the OXTR antagonist i.p.-injected rats. These results suggest that central oxytocinergic pathways may be involved in anti-inflammation at the spinal level and suppression of feeding behavior at the forebrain-brainstem level in AA rats.
Angioni L, Cocco C, Ferri G, Argiolas A, Melis M, Sanna F (2016) Involvement of nigral oxytocin in locomotor activity: A behavioral, immunohistochemical and lesion study in male rats. Horm Behav 83:23-38. doi: 10.1016/j.yhbeh.2016.05.012
Summary: Oxytocin is well known for its hormonal role in lactation and parturition, but also exerts widespread actions in central nervous system. Previous experiments revealed the existence of a correlation between the changes in locomotor activity found in Oxytocin-SAP-treated rats and the extent of the changes in nigral TH and vesicular glutamate transporters immunoreactivity, provide support for a modulatory role of oxytocin on locomotor activity at the level of the substantia nigra. The day after a prior assessment of spontaneous locomotor activity, rats were randomly injected bilaterally with 0.3 μL of Oxytocin-SAP (Cat. #IT-46, 60 ng/μL/site), or with the same amount of Blank-SAP (Cat. #IT-21, 60 ng/μL/site) or with vehicle (0.3 μL/site of PBS, pH 7.4). Whether oxytocin may be considered as a target for controlling motor disturbances, as those occurring in Parkinson’s disease and/or in other motor disturbances related to basal ganglia dysfunctions, remains to be evaluated
Mittelman-Smith M, Krajewski-Hall S, McMullen N, Rance N (2016) Ablation of KNDy neurons results in hypogonadotropic hypogonadism and amplifies the steroid-induced LH surge in female rats. Endocrinology 157:2015-2027. doi: 10.1210/en.2015-1740
Summary: KNDy neurons are a subpopulation of neurons in the infundibular nucleus that coexpress estrogen receptor α, kisspeptin, and neurokinin B (NKB) mRNA. Previous work indicated that altered signaling from KNDy neurons may play a role in the low levels of circulating sex steroids found in hypogonadotropic hypogonadism. Rats received bilateral 10-ng injections of NK3-SAP (Cat. #IT-63) dorsal to the arcuate nucleus. Blank-SAP (Cat. #IT-21) was used as control. In animals with intact ovaries the NK3-SAP lesion resulted in hypogonadotropic hypogonadism. In contrast, the LH surge in lesioned ovariectomized rats was 3-fold higher, demonstrating that KNDy neurons are integral for the control of serum LH levels, estrous cyclicity, and may also have some control over the magnitude of the LH surge.
Fu R, Chen X, Zuo W, Li J, Kang S, Zhou L, Siegel A, Bekker A, Ye J (2016) Ablation of μ opioid receptor-expressing GABA neurons in rostromedial tegmental nucleus increases ethanol consumption and regulates ethanol-related behaviors. Neuropharmacology 107:58-67. doi: 10.1016/j.neuropharm.2016.02.027
Summary: In this work the authors investigated cellular mechanisms underlying the aversive effects of alcohol that limit its intake. Previous work has linked synaptic inhibition of dopamine neurons in the ventral tegmental area to this aversion. Rats conditioned to ingest ethanol received bilateral injections totaling 3 pmol of Dermorphin-SAP (Cat. #IT-12) into the rostromedial tegemental nucleus (RTMg). Blank-SAP (Cat. #IT-21) was used as a control. Lesioned animals displayed significantly increased preference for, and intake of ethanol, while showing no change in the desire for sucrose. The results indicate that mu opioid expressing GABAergic neurons in the RTMg are highly involved in the regulation of ethanol consumption.
Li P, Janczewski W, Yackle K, Kam K, Pagliardini S, Krasnow M, Feldman J (2016) The peptidergic control circuit for sighing. Nature 530:293-297. doi: 10.1038/nature16964
Summary: Sighs are often associated with relief or sadness, but rodents sigh spontaneously dozens of times per hour. There are physiological benefits to sighing, including enhancement of gas exchange and preservation of lung integrity. The authors identify a peptidergic sigh control circuit in the retrotrapezoid nucleus/parafacial respiratory group of the mouse brain that projects to the pre-Bötzinger complex. Mice received bilateral 6.2-ng injections of Bombesin-SAP (Cat. #IT-40) into the pre-Bötzinger complex. Blank-SAP (Cat. #IT-21) was used as control. Elimination of the bombesin receptor-expressing neurons or inhibition of neuromedin B receptor-expressing neurons suppressed sighing. Interfering with the activity of both receptors abolished sigh activity while leaving normal breathing intact. The data suggest that overlapping peptidergic pathways are the core of a sigh control circuit.
Hulsey D, Hays S, Khodaparast N, Ruiz A, Das P, Rennaker R, Kilgard M (2016) Reorganization of motor cortex by vagus nerve stimulation requires cholinergic innervation. Brain Stimul 9:174-181. doi: 10.1016/j.brs.2015.12.007
Summary: Recent work has suggested that vagus nerve stimulation (VNS) can enhance neuroplasticity, and coupled with other training can drive motor cortex reorganization. These findings highlight the potential of VNS to support recovery from neurological disease. Pretrained rats received bilateral injections totaling 3.75 μg of 192-IgG-SAP (Cat. #IT-01) into the nucleus basalis (NB). Mouse-IgG-SAP (Cat. #IT-18) was used as control. Control animals displayed a substantial increase in proximal limb representation, lesion of the NB prevented this increase. Motor performance was similar between lesion and control groups, indicating that the difference in representation was not due to altered limb function.
Arora V, Morado-Urbina C, Aschenbrenner C, Hayashida K, Wang F, Martin T, Eisenach J, Peters C (2016) Disruption of spinal noradrenergic activation delays recovery of acute incision-induced hypersensitivity and increases spinal glial activation in the rat. J Pain 17:190-202. doi: 10.1016/j.jpain.2015.10.009
Summary: A significant percentage of patients who undergo surgery experience prolonged clinically impactful pain, reducing the quality of life and physical function. Disruption of the descending noradrenergic input has been hypothesized to be important to the generation of this type of pain state. Using an acute incision model, the authors administered 5 μg ofAnti-DBH-SAP (Cat. #IT-03) to the L5-L6 interspace of rats. Mouse IgG-SAP (Cat. #IT-18) was used as a control. Lesioned animals demonstrated a significant increase in mechanical hypersensitivity, and a smaller increase in thermal hypersensitivity. This and other results suggest that spinally projecting noradrenergic pathways are necessary for normal recovery from surgical incision, and possibly other types of pain.
Freiria-Oliveira A, Blanch G, Pedrino G, Cravo S, Murphy D, Menani J, Colombari D (2015) Catecholaminergic neurons in the comissural region of the nucleus of the solitary tract modulate hyperosmolality-induced responses. Am J Physiol Regul Integr Comp Physiol 309:R1082-1091. doi: 10.1152/ajpregu.00432.2014
Summary: Body fluid homeostasis and cardiovascular regulation are thought to be at least in part controlled by noradrenergic A2 neurons found in the nucleus of the solitary tract (NTS). In this work the authors investigated the involvement of A2 neurons of the commissural NTS in arterial pressure, as well as several body fluid homeostasis parameters. Rats received 12.6-ng injections of Anti-DBH-SAP (Cat. #IT-03) into the commissural NTS. Mouse IgG-SAP (Cat. #IT-18) was used as a control. Lesioned animals displayed increased c-Fos expression in the hypothalamic paraventricular nucleus when treated with hypertonic NaCl, and increased arterial pressure. The data indicate that commissural NTS A2 neurons are essential for inhibitory mechanisms that reduce water intake and pressor response to an acute increase in plasma osmolality.
Wiley RG (2015) Nociceptive effects of neurotensin(NTS)- and somatostatin(SST)-toxin conjugates applied to the lumbar dorsal horn in rats. Neuroscience 2015 Abstracts 418.11/O12. Society for Neuroscience, Chicago IL.
Summary: Intrathecal injections of NTS or SST have been reported to be anti-nociceptive, and in the case of SST, analgesic in humans. Preliminary experiments in our lab previously showed that lumbar intrathecal injection of the excitatory neuropeptide, NTS, or the inhibitory neuropeptide, SST, conjugated to the ribosome inactivating protein, saporin (sap), produced compulsive scratching/biting of hindquarters resulting in loss of fur and skin. This was thought likely due to pain and/or itching from selective loss of superficial dorsal horn nociceptive inhibitory interneurons expressing NTS receptors. Subsequent experiments using lumbar intrathecal injections of NTS-cholera toxin A chain conjugate resulted in prolonged anti-nociception on hotplate, tail flick and von Frey testing, that was not reversed by naloxone and lasted several days, likely due to sustained activation of the same neurons. The present study sought to determine if the lesions produced by NTS-sap or SST-sap alter nociceptive responses. In the present study, rats, under isoflurane anesthesia, were injected intrathecally using temporarily-placed subarachnoid catheters over the lumbar enlargement with 10 ul of sterile preservative-free normal saline containing either 300-400 ng of NTS-sap, 1 ug of SST-sap or 1 ug blank-sap (control) from Advanced Targeting Systems, San Diego, CA. Catheters were flushed with an additional 10 ul of saline. After post-surgical recovery, the rats were then observed for scratching/biting their hindquarters, nocifensive responses on the hotplate, von Frey mechanical probing of the hindpaws, and on operant thermal escape. 4 of 11 NTS-saporin rats and 5 of 9 SST-saporin rats, but none of 9 blank-saporin rats began scratching within 8-47 days after toxin conjugate injection. Hotplate nocifensive reflex testing at 44.5°C and 47°C showed no significant difference between the groups. Von Frey, operant thermal escape testing and anatomic studies are in progress to further specify the functional effects of the toxin conjugate injections and to identify the dorsal horn neurons being destroyed. The results to date are interpreted as consistent with a possibly unique role for NTS and/or SST receptor-expressing superficial dorsal horn inhibitory interneurons in nociception and/or itch. Excitatory/activating moieties such as cholera toxin A subunit targeted by conjugation to NTS or SST may offer a novel approach to enhance inhibition in nociceptive dorsal horn neurons and to produce analgesia by a non-opioid mechanism.
Zoccal DB, Taxini CL, Gargaglioni LH (2015) Control of sympathetic activity by A5 noradrenergic neurons in the in situ rat preparations. Neuroscience 2015 Abstracts 432.16/X11. Society for Neuroscience, Chicago IL.
Summary: The A5 area represents an important noradrenergic neuronal group located in the ventral pons that receives and sends projections to various medullary areas involved in the cardiorespiratory control. Its involvement in the chemoreflex control was previously studied in anesthetized conditions. In the present study, we explored the contribution of A5 noradrenergic neurons in the processing of sympathetic responses to central and peripheral chemoreceptors stimulation using the in situ working heart-brainstem rat preparation. Juvenile male Holztman rats received bilateral microinjections of either IgG-SAP (50nl, n=7) or toxin anti-dopamine beta-hydroxylase-saporin (anti-DβH-SAP, 4.2 ng/50 nl, n=6) in the A5. One week later, in situ preparations were obtained to record the thoracic sympathetic (tSN) and phrenic nerve (PN) activities; and stimulation of peripheral (KCN, 0.05%, 50nL) and central chemoreceptors (7 and 10% CO2 in the perfusate, 5 min) were performed. Baseline tSN activity (12.5±2.0 vs 12.6±2.4 μV), PN burst amplitude (40.7±9.7 vs 44.8 ±19.9 μV) and frequency (13±1 vs 15±2 bpm) and the respiratory-sympathetic coupling pattern were similar between control and A5-lesioned rats. The sympathetic ([[unable to display character: ∆]]tSN: 110±12 vs 58±8 %, P<0.05), but not the phrenic response to peripheral chemoreflex stimulation was marked attenuated in animals with lesion of A5 noradrenergic neurons. As to the central chemoreflex, the tSN response to 7% CO2 tSN: 9.5±1.4 vs 3.9±1.7%, P<0.05), but not to 10% CO2 (16.4±2.9 vs 10.9±1.6%) was lower in A5-lesioned rats in comparison to controls. On the other hand, the PN response to 7 and 10% CO2 were similar between control and A5-lesioned rats. Our data show that the A5 noradrenergic neurons are critical for the full expression of the sympathetic chemoreflex responses, possibly by providing an excitatory drive to the neurons generating sympathetic activity.
Pittenger CJ (2015) Modeling Tourette syndrome pathophysiology through targeted manipulation of striatal interneurons. Neuroscience 2015 Abstracts 6.07. Society for Neuroscience, Chicago IL.
Summary: Postmortem studies of Tourette syndrome patients has revealed a reduction in the number of specific striatal interneurons. The authors explored the hypothesis that this neuronal deficit is enough to produce the symptoms of Tourette syndrome in mice. Animals received 90-ng injections of Anti-ChAT-SAP (Cat. #IT-42) into the striatum. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data suggest that loss of the striatal interneurons is enough to produce some, but not all, of the symptoms caused by Tourette syndrome.
Pires E, D'Souza R, Needham M, Herr A, Jazaeri A, Li H, Stoler M, Anderson-Knapp K, Thomas T, Mandal A, Gougeon A, Flickinger C, Bruns D, Pollok B, Herr J (2015) Membrane associated cancer-oocyte neoantigen SAS1B/ovastacin is a candidate immunotherapeutic target for uterine tumors. Oncotarget 6:30194-30211. doi: 10.18632/oncotarget.4734
Summary: Ovastatin is a zinc matrix metallo-proteinase thought to play roles in sperm-egg interaction and the prevention of polyspermy in eutherians. This protein is not found in normal adult tissues, but is expressed by uterine carcinosarcomas. The authors investigated the possibility of targeting ovastatin as a tumor surface neoantigen for therapeutic purposes. SNU539 cells, a uterine malignant mixed Müllerian tumor-derived cell line, were challenged with 1 μM, 0.1 μM, and 0.01 μM rabbit polyclonal anti-ovastatin coupled to 5.42 nM Fab-ZAP rabbit (Cat. #IT-57). Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The results indicate that for this form of uterine cancer, ovastatin is a viable therapeutic target.
Devoto P, Flore G, Saba P, Frau R, Gessa G (2015) Selective inhibition of dopamine-beta-hydroxylase enhances dopamine release from noradrenergic terminals in the medial prefrontal cortex. Brain Behav 5:e00393. doi: 10.1002/brb3.393
Summary: Dopamine-beta-hydroxylase (DBH) is a neuronal enzyme that is a potential target for the treatment of cocaine abuse, alcohol dependence, and eating disorders. Here the authors administered 5 μg of icv Anti-DBH-SAP (Cat. #IT-03) to rats, and assessed the effect of the dopaminergic lesion on levels of extracellular dopamine. Mouse IgG-SAP (Cat. #IT-18) and saporin (Cat. #PR-01) were used as controls. Extracellular levels of dopamine were significantly increased in both lesioned animals and those treated with the DBH inhibitor nepicastat. Clonadine could reverse the nepicastat effect, but not the effect of Anti-DBH-SAP treatement. The data demonstrate a mechanism for the synergistic effect of cocaine on nepicastat-induced dopamine release.
Cao Q, Lu J, Li Q, Wang C, Wang X, Lee V, Wang C, Nguyen H, Zheng G, Zhao Y, Alexander S, Wang Y, Harris D (2016) CD103+ dendritic cells elicit CD8+ t cell responses to accelerate kidney injury in adriamycin nephropathy. J Am Soc Nephrol 27:1344-1360. doi: 10.1681/ASN.2015030229
Summary: Although it is known that dendritic cells (DCs) are involved in chronic kidney disease, it is not well understood how they either resolve or aggravate the condition. CD103+ dendritic cells in particular, are known to maintain tolerance through interaction with regulatory T cells, as well as protect against infection through interactions with CD8+ T cells. In this work the authors depleted CD103+ DCs by administering 1 mg/kg of anti-CD103-SAP (Cat. #IT-50) to the intraperitoneal space of mice subject to adriamycin nephropathy. Rat IgG-SAP (Cat. #IT-17) was used as a control. Elimination of the CD103+ DCs attenuated the kidney injury, indicating that in murine chronic kidney disease CD103+ DCs are pathogenic rather than therapeutic.
Oyamaguchi A, Abe T, Sugiyo S, Niwa H, Takemura M (2016) Selective elimination of isolectin B4-binding trigeminal neurons enhanced formalin-induced nocifensive behavior in the upper lip of rats and c-Fos expression in the trigeminal subnucleus caudalis. Neurosci Res 103:40-47. doi: 10.1016/j.neures.2015.07.007
Summary: In adult rats non-peptidergic neurons and peptidergic neurons innervate different areas and layers of the lamina. It is thought that these two neuronal populations play different roles in nociceptive processing, but the specific function of each group is not well understood. In order to investigate peptidergic and non-peptidergic neurons in orofacial pain processing the authors injected the cisterna magna of rats with 2.9 μg of rIB4-SAP (Cat. #IT-10). Blank-SAP (Cat. #IT-21) was used as a control. The lesioned animals displayed more frequent face-rubbing responses on the administration of formalin, indicating that IB4-binding neurons in the trigeminal nerve play an anti-nociceptive role in response to this type of pain.
King T, Ruyle B, Kline D, Heesch C, Hasser E (2015) Catecholaminergic neurons projecting to the paraventricular nucleus of the hypothalamus are essential for cardiorespiratory adjustments to hypoxia. Am J Physiol Regul Integr Comp Physiol 309:R721-731. doi: 10.1152/ajpregu.00540.2014
Summary: Catecholaminergic neurons in the brainstem are known to be involved in cardiorespiratory control and to modulate sensory function. Some of the projections from these neurons are to the paraventricular nucleus (PVN), and are involved in cardiorespiratory and neuroendocrine responses to hypoxia. While data have shown the PVN-projecting neurons are activated by hypoxia, their function in this context is not known. In this work the authors bilaterally injected 42 ng of Anti-DBH-SAP (Cat. #IT-03) into the PVN of rats. Mouse IgG-SAP (Cat. #IT-18) was used as control. Respiratory measurements of the lesioned animals indicates that PVN-projecting catecholaminergic neurons are involved in peripheral and central chemoreflex and arterial oxygen levels during exposure to hypoxic stimuli.
Akiyama T, Nguyen T, Curtis E, Nishida K, Devireddy J, Delahanty J, Carstens M, Carstens E (2015) A central role for spinal dorsal horn neurons that express neurokinin-1 receptors in chronic itch. Pain 156:1240-1246. doi: 10.1097/j.pain.0000000000000172
Summary: Chronic itch is caused by increased sensitivity of itch-signaling pathways. It can be generated by normally itchy stimuli (hyperknesis) and by normally non-itchy light touch (alloknesis). The authors used an ovalbumin-induced atopic dermatitis model to study chronic itch in mice. The mice received 400-ng intrathecal injections of Bombesin-SAP (Cat. #IT-40), SSP-SAP (Cat. #IT-11), or the control Blank-SAP (Cat. #IT-21). While Bombesin-SAP significantly attenuated hyperknesis, it had no effect on spontaneous scratching or alloknesis. SSP-SAP reduced all behavioral signs of chronic itch.
Bourassa E, Stedenfeld K, Sved A, Speth R (2015) Selective C1 lesioning slightly decreases angiotensin II type I receptor expression in the rat rostral ventrolateral medulla (RVLM). Neurochem Res 40:2113-2120. doi: 10.1007/s11064-015-1649-3
Summary: Exogenous angiotensin II administered to the RVLM produces a significant pressor response that can be countered by angiotensin II type I receptor antagonists. In this work the authors examined the relative contribution of C1 and non-C1 neurons in the RVLM to this angiotensin II response. Rats received 10 or 15 ng of Anti-DBH-SAP (Cat. #IT-03) as unilateral injections into the RVLM. Mouse IgG-SAP (Cat. #IT-18) was used as control. The data indicate that the majority of angiotensin II type 1 receptors are expressed on non-C1 neurons or glia.
Peters C, Hayashida K, Suto T, Houle T, Aschenbrenner C, Martin T, Eisenach J (2015) Individual differences in acute pain-induced endogenous analgesia predict time to resolution of postoperative pain in the rat. Anesthesiology 122:895-907. doi: 10.1097/ALN.0000000000000593
Summary: The authors investigated the relationship between preoperative Conditioned Pain Modulation (CPM) and the time course of recovery from surgery. CPM was evaluated using forepaw capsaicin injections into rats. During the study, lesioned rats received 5-μg intrathecal injections of anti-DBH-SAP (Cat. #IT-03), followed 14 days later by a partial L5 spinal nerve ligation surgery. Mouse-IgG-SAP (Cat. #IT-18) was used as a control. CPM was partially blocked in the lesioned animals, suggesting descending noradrenergic signaling is important in the time course of recovery from surgery.
Mittelman-Smith M, Krajewski-Hall S, McMullen N, Rance N (2015) Neurokinin 3 receptor-expressing neurons in the median preoptic nucleus modulate heat-dissipation effectors in the female rat. Endocrinology 156:2552-2562. doi: 10.1210/en.2014-1974
Summary: Kisspeptin and Neurokinin B (NKB) expression in the infundibular, or arcuate, nucleus is increased after menopause. Here the authors investigate whether KNDy (kisspeptin, NKB, and dynorphin expressing) neurons are able to influence cutaneous vasodilation through Neurokinin 3 (NK3)-expressing projections from the median preoptic nucleus (MnPO). Rats received two 10-ng injections of NK3-SAP (Cat. #IT-63) into the MnPO. Blank-SAP (Cat. #IT-21) was used as a control. The data indicate that NK3-expressing neurons in the MnPO facilitate vasodilation.
Xu M, Kobets A, Du J, Lennington J, Li L, Banasr M, Duman R, Vaccarino F, DiLeone R, Pittenger C (2015) Targeted ablation of cholinergic interneurons in the dorsolateral striatum produces behavioral manifestations of Tourette syndrome. Proc Natl Acad Sci U S A 112:893-898. doi: 10.1073/pnas.1419533112
Summary: Postmortem studies of Tourette syndrome patients has revealed a reduction in the number of specific striatal interneurons. The authors explored the hypothesis that this neuronal deficit is enough to produce the symptoms of Tourette syndrome in mice. Animals received 90-ng injections of Anti-ChAT-SAP (Cat. #IT-42) into the striatum. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data suggest that loss of the striatal interneurons is enough to produce some, but not all, of the symptoms caused by Tourette syndrome.
Cetas J, McFarlane R, Kronfeld K, Smitasin P, Liu J, Raskin J (2015) Brainstem opioidergic system is involved in early response to experimental SAH. Transl Stroke Res 6:140-147. doi: 10.1007/s12975-014-0378-2
Objective: To determine the cause of poor long-term outcomes after Subarachnoid hemorrhage (SAH).
Summary: Failure of the RVM μ-opioid receptor cells to initiate the compensatory CBF response sets the stage for acute and delayed ischemic injury following SAH.
Usage: To lesion medullary neurons expressing the μ-opioid receptor, Dermorphin–SAP was microinjected as a bilateral dose of 0.5 pmol in 500 nL per side (1 pmol in 1 μL total dose and injection volume). Blank–SAP or vehicle was injected in equal volumes and dose as controls.
Zhao Z, Wan L, Liu X, Huo F, Li H, Barry D, Krieger S, Kim S, Liu Z, Xu J, Rogers B, Li Y, Chen Z (2014) Cross-inhibition of NMBR and GRPR signaling maintains normal histaminergic itch transmission. J Neurosci 34:12402-12414. doi: 10.1523/JNEUROSCI.1709-14.2014
Summary: After itch detection, the itch pathway moves through an array of G-protein coupled receptors and transient receptor potential channels in dorsal root ganglion neurons into dorsal horn neurons which integrate and transduce these signals, sending them to the somatosensory cortex. The purpose of this work is to clarify whether gastrin-releasing peptide (GRP) or B-type natriuretic peptide regulates histaminergic itch. Several strains of knockout mice received 200, 300, or 400 ng intrathecal injections of bombesin-SAP (Cat. #IT-40). Blank-SAP (Cat. #IT-21) was used as a control. The data further define the respective functions of the neuromedin B receptor and GRP receptor in itch, and reveals a working relationship between the different interneuron populations.
Lee CL, Rajkumar R, Dawe GS (2014) Featured Article: Corticotropin releasing factor-saporin conjugate selectively lesions nucleus incertus. Targeting Trends 15(2)
Jokiaho A, Donovan C, Watts A (2014) The rate of fall of blood glucose determines the necessity of forebrain-projecting catecholaminergic neurons for male rat sympathoadrenal responses. Diabetes 63:2854-2865. doi: 10.2337/db13-1753
Summary: Different sets of glucosensors detect insulin-induced hypoglycemia depending on the onset rate. This detection controls the activation of sympathoadrenal counterregulatory responses (CRRs). Slow onset hypoglycemia, common with insulin therapy, is detected by glucosensors in the portal-mesenteric veins. Fast onset is detected by brain elements. The authors lesioned hindbrain catecholaminergic neurons to determine which set of responses-they interact with. Rats received 42 ng bilateral injections of Anti-DBH-SAP (Cat. #IT-03) into the paraventricular nucleus of the hypothalamus. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data indicate that these neurons are critical for detection of slow-onset insulin-induced hypoglycemia.
Ogawa S, Nathan FM, Parhar IS (2014) Habenular kisspeptin modulates fear in the zebrafish. Proc Natl Acad Sci U S A 111(10):3841-3846. doi: 10.1073/pnas.1314184111
Summary: The peptide kisspeptin can be found in several areas of the brain, but its role in regions other than the hypothalamus has not been studied. Zebrafish express kiss1 mRNA which is a conserved ortholog of the mammalian KISSI/KissI making zebrafish a viable model for investigating the role of kisspeptin in various brain systems. Animals received 1 μg of the custom conjugate kiss-SAP (see NK3-SAP, Cat. #IT-63) via an intracranial injection. Blank-SAP (Cat. #IT-21) was used as a control. Reducing Kiss1 immunoreactivity in the habenula and the raphe reduced an invoked fear response, indicating a role for kisspeptin in fear inhibition.
Lee SC, Seo KW, Kim HJ, Kang SW, Choi HJ, Kim A, Kwon BS, Cho HR, Kwon B (2015) Depletion of alloreactive T cells by anti-CD137-saporin immunotoxin. Cell Transplant 24:1167-1181. doi: 10.3727/096368914X679327
Summary: The ability to selectively remove T cells that mediate graft-versus-host disease (GVHD) would prevent graft rejection as well as preserve the T cells that mediate graft-versus-leukemia (GVL) effect – especially in cases of hematopoietic stem cell transplantation. In this work the authors used a custom conjugate of anti-mouse CD137 and saporin to eliminate alloreactive T cells during a T cell donor transfer in mice. Rat IgG-SAP (Cat. #IT-17) was used as a control. Transfer of T cells after the immunotoxin treatment did not cause GVHD, but the GVL effect was left intact, indicating the potential of selective T cell depletion for transplantation.
Related Products: Rat IgG-SAP (Cat. #IT-17)
Suto T, Severino AL, Eisenach JC, Hayashida KI (2014) Gabapentin increases extracellular glutamatergic level in the locus coeruleus via astroglial glutamate transporter-dependent mechanisms. Neuropharmacology 81C:95-100. doi: 10.1016/j.neuropharm.2014.01.040
Summary: Gabapentin is effective in reducing acute and chronic pain, but the mechanisms by which it works are not well understood. The authors assessed extracellular glutamate levels and glutamate interaction with several different cellular membrane proteins. Rats received a 0.25 μg injection of anti-DBH-SAP (Cat. #IT-03) into the locus coeruleus (LC) in order to deplete noradreline levels. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The gabapentin-induced glutamate increase in the LC was not affected by the lesion, supporting data indicating that gabapentin induces glutamate release from astrocytes to stimulate descending inhibition.
Lee LC, Rajkumar R, Dawe GS (2014) Selective lesioning of nucleus incertus with corticotropin releasing factor-saporin conjugate. Brain Res 1543:179-190. doi: 10.1016/j.brainres.2013.11.021
Summary: In this work the authors used CRF-SAP (Cat. #IT-13) to eliminate CRF1 receptor-expressing cells from the nucleus incertus (NI). Rats received bilateral CRF-SAP injections of 21.5 to 86 ng into the NI. Blank-SAP (Cat. #IT-21) was used as a control. Lesioned animals displayed a significant loss of CRF1 receptor-expressing cells, along with a decrease in relaxin-3 and GAD65 expression.
Weisshaar CL, Winkelstein BA (2014) Ablating spinal NK1-bearing neurons eliminates the development of pain and reduces spinal neuronal hyperexcitability and inflammation from mechanical joint injury in the rat. J Pain 15(4):378-386. doi: 10.1016/j.jpain.2013.12.003
Summary: A high percentage of chronic neck pain involves the facet joint. Although the facet joint is innvervated by peptide-responsive nociceptive afferents, the role of these cells in the development and modulation of nociceptive signaling remains unclear. Using a previously developed rat model of facet joint injury, the authors examined the role of neurokinin-1 receptor-expressing spinal cells in this pathway. Rats received 100 ng SSP-SAP (Cat. #IT-11) via lumbar puncture. Blank-SAP (Cat. #IT-21) was used as a control. The results demonstrate that spinal NK1r-expressing cells are essential for nociception and inflammation due to a mechanical joint injury.
Freiria-Oliveira AH, Blanch GT, De Paula PM, Menani JV, Colombari DS (2013) Lesion of the commissural nucleus of the solitary tract/A2 noradrenergic neurons facilitates the activation of angiotensinergic mechanisms in response to hemorrhage. Neuroscience 254:196-204. doi: 10.1016/j.neuroscience.2013.09.017
Summary: Previous work has generated conflicting data on the role of catecholaminergic A2 neurons in the nucleus of the solitary tract (NTS) in control of arterial pressure lability. The authors used Anti-DBH-SAP (Cat. #IT-03) to lesion these neurons in a hypotensive hemorrhage model. Rats received two injections of 12.6 ng into the commissural NTS. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The lesioned animals quickly recovered from hypotension, but were impaired by the icv administration of losartan.
Romano A, Potes CS, Tempesta B, Cassano T, Cuomo V, Lutz T, Gaetani S (2013) Hindbrain noradrenergic input to the hypothalamic PVN mediates the activation of oxytocinergic neurons induced by the satiety factor oleoylethanolamide. Am J Physiol Endocrinol Metab 305(10):E1266-73. doi: 10.1152/ajpendo.00411.2013
Summary: Feeding behavior and energy balance are in part controlled by signals from the gut. Oleoylethanolamide (OEA) is an acylethanolamide that is thought to play a role in this network. Since peripheral administration of OEA has effects on the nucleus of the solitary tract (NTS) and paraventricular nucleus (PVN) the authors investigated the role of noradrenergic afferent input to these areas. Rats received bilateral 84-ng injections of Anti-DBH-SAP (Cat. #IT-03) into the PVN. Mouse IgG-SAP (Cat. #IT-18) was used as a control.
Laplante F, Dufresne MM, Ouboudinar J, Ochoa-Sanchez R, Sullivan RM (2013) Reduction in cholinergic interneuron density in the nucleus accumbens attenuates local extracellular dopamine release in response to stress or amphetamine. Synapse 67(1):21-29. doi: 10.1002/syn.21612
Summary: The authors examined whether excessive dopamine neurotransmission in the mesolimbic system is due to higher levels of presynaptic or postsynaptic dopamine. Rats received 250-ng bilateral injections of anti-ChAT-SAP (Cat. #IT-42) into the nucleus accumbens. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data suggest that reduction of cholinergic interneurons in the nucleus accumbens suppresses presynaptic dopamine release.
Wiater MF, Li AJ, Dinh TT, Jansen HT, Ritter S (2013) Leptin-sensitive neurons in the arcuate nucleus integrate activity and temperature circadian rhythms and anticipatory responses to food restriction. Am J Physiol Regul Integr Comp Physiol 305(8):R949-R960. doi: 10.1152/ajpregu.00032.2013
Summary: The arcuate nucleus (Arc) of the hypothalamus is known to participate in the regulation of feeding, adiposity, and leptin-dependent metabolism. The authors examined the role of leptin-receptive neurons in locomotor and temperature rhythms. Rats received four bilateral injections of Leptin-SAP (Cat. #IT-47) into the Arc; Blank-SAP (Cat. #IT-21) was used as a control. The lesion affected learning connected to light cycles, but not learning connected to food schedules, suggesting a mechanism for internal desynchrony that might play a role in obesity and other metabolic disorders.
Khasabov SG, Simone DA (2013) Loss of neurons in rostral ventromedial medulla that express neurokinin-1 receptors decreases the development of hyperalgesia. Neuroscience 250C:151-165. doi: 10.1016/j.neuroscience.2013.06.057
Summary: Previous data has indicated that neurokinin-1 receptors are located on ON cells in the rostral ventromedial medulla (RVM). ON cells are considered pronociceptive because noxious stimulation is stimulatory. In this work the authors eliminated ON cells using 0.3-μl injections of 1 μM SSP-SAP (Cat. #IT-11) into the left and right side of the RVM. Blank-SAP (Cat. #IT-21) was used as a control. SSP-SAP treatment did not change mechanical or heat withdrawal responses, or change morphine-induced analgesia. A significant reduction in the duration of nocifensive behaviors induced by various hyperalgesic stimulators indicated that these neurons are involved in pain facilitation rather than modulation.
Sasaki A, Adhikari S, Andoh T, Kuraishi Y (2013) BB2 bombesin receptor-expressing spinal neurons transmit herpes-associated itch by BB2 receptor-independent signaling. Neuroreport 24(12):652-656. doi: 10.1097/WNR.0b013e32836352d8
Summary: Using a skin rash model created by inoculating mice with human herpes virus, bombesin receptor-expressing spinal neurons were lesioned intrathecally with 400 ng of Bombesin-SAP (Cat. #IT-40). Lesioned animals displayed reduced scratching, but licking (due to pain) was not reduced.
Helena CV, Kalil B, Anselmo-Franci JA, Bertram R (2013) Time course study of targeted ablation of KNDy neurons, but not tyrosine-hydroxylase neurons, in the rat arcuate nucleus using a neurokinin b-saporin. Endocr Rev 34:OR47-5. 95th Annual Meetin and Expo, San Francisco. doi: 10.1093/edrv/34.supp.1
Summary: Ovariectomized rats were given bilateral injections of NK3-SAP (Cat. #IT-63) into the arcuate nucleus for a time course study of KNDy neuron loss. Blank-SAP (Cat. #IT-21) was used as a control.
Fricks-Gleason AN, Keefe KA (2013) Evaluating the role of neuronal nitric oxide synthase-containing striatal interneurons in methamphetamine-induced dopamine neurotoxicity. Neurotox Res 24(2):288-297. doi: 10.1007/s12640-013-9391-6
Summary: Using the fact that neuronal nitric oxide synthase (nNOS)-containing neurons in the striatum express the substance P receptor, the authors injected four locations in the striatum with 3 ng each of SSP-SAP (Cat. #IT-11). Blank-SAP (Cat. #IT-21) was used as a control. Although there was a significant loss of nNOS-containing neurons, the lesions did not attenuate NO production.
Hamlin AS, Windels F, Boskovic Z, Sah P, Coulson EJ (2013) Lesions of the basal forebrain cholinergic system in mice disrupt idiothetic navigation. PLoS One 8(1):e53472. doi: 10.1371/journal.pone.0053472
Summary: Alzheimer's disease patients perform poorly on spatial navigation tests requiring either distal cues (allothetic) or body-centered cues (idiothetic). The authors used 0.2 μg bilateral infusions of mu p75-SAP (Cat. #IT-16) into the lateral ventricles of mice to examine the hypothesis that the cholinergic medial septo-hippocampal circuit is important for idiothetic navigation. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. Lesioned animals were similar to controls in contextual fear conditioning, spatial working memory, as well as several other parameters. But exploratory behavior requiring idiothetic signals was very disorganized, indicating that cholinergic cells are vital to idiothetic navigation.
LaPlante F (2013) Featured Article: Role of cholinergic neurons in the nucleus accumbens and their involvement in schizophrenic pathology. Targeting Trends 14(1)
Intact catecholamine inputs to the forebrain are required for appropriate regulation of corticotrophin-releasing hormone and vasopressin gene expression by corticosterone in the rat paraventricular nucleus.
Kaminski KL, Watts AG (2012) Intact catecholamine inputs to the forebrain are required for appropriate regulation of corticotrophin-releasing hormone and vasopressin gene expression by corticosterone in the rat paraventricular nucleus. J Neuroendocrinol 24(12):1517-1526. doi: 10.1111/j.1365-2826.2012.02363.x
Summary: Corticosterone releasing hormone (CRH) neurons in the paraventricular nucleus of the hypothalamus (PVH) control release of adrenocorticotropic hormone and glucocorticoids. In order to determine the contribution of these neurons to CRH and vasopressin expression in the PVH the authors administered bilateral 42 ng injections of anti-DBH-SAP (Cat. #IT-03) into the PVH of both normal and adrenalectomized rats. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data demonstrate that under certain conditions CRH and vasopressin gene expression is modulated by interactions between corticosterone and catecholaminergic projections to the hypothalamus.
Cusulin C, Monni E, Ahlenius H, Wood J, Brune J, Lindvall O, Kokaia Z (2012) Embryonic stem cell-derived neural stem cells fuse with microglia and mature neurons. Stem Cells 30:2657-2671. doi: 10.1002/stem.1227
Summary: The fusogenic role of microglia could be even more important after NSC transplantation into brains affected by neurodegenerative diseases associated with microglia activation.
Usage: Primary Cells and NS Cell Coculture. Seven to twelve days after plating primary cells, NS cells were plated on top (10,000 cells per cm2) for 1–3 days. Rat primary cells were treated with 10 nM Mac-1-SAP or Mouse IgG-SAP during the 5 days prior to the coculture, and analyzed 3 days thereafter.
Kato J, O’Donnell RT, Abuhay M, Tuscano JM (2012) Efficacy and toxicity of a CD22-targeted antibody-saporin conjugate in a xenograft model of non-Hodgkin’s lymphoma. Oncoimmunology 1(9):1469-1475. doi: 10.4161/onci.21815
Summary: CD22 is a B-cell-specific antigen found on many B-cell malignancies. It is not expressed by stem cell precursors, and is rapidly internalized when bound by an antibody. In this work, the authors use a custom conjugate of anti-CD22 (mAb HB22.7) and saporin in a cytotoxicity assay on non-Hodgkin's lymphoma cell lines, as well as in a mouse tumor model. The dosing for the tumor model was 1 mg conjugate per kg of animal. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The results indicate that CD22 is a potential therapeutic target for cancer therapy.
Mittelman-Smith MA, Williams H, Krajewski-Hall SJ, McMullen NT, Rance NE (2012) Role for kisspeptin/neurokinin B/dynorphin (KNDy) neurons in cutaneous vasodilatation and the estrogen modulation of body temperature. Proc Natl Acad Sci U S A 109(48):19846-19851. doi: 10.1073/pnas.1211517109
Summary: Menopause is marked by estrogen withdrawal, and also by hot flushes. Given the fact that hypothalamic levels of kisspeptin/neurokinin B/dynorphin (KNDy) neurons are significantly altered in menopause, the authors investigated whether these neurons are involved in the generation of flushes. Rats received bilateral injections of NK3-SAP (Cat. #IT-63) into the arcuate nucleus – a total of 40 ng. Blank-SAP (Cat. #IT-21) was used as control. The data indicate that KNDy neurons promote cutaneous vasodilation, and play a role in 17β-estradiol modulation of body temperature, supporting the hypothesis that these neurons could play a role in the generation of hot flushes.
Kerbler GM, Hamlin AS, Pannek K, Kurniawan ND, Keller MD, Rose SE, Coulson EJ (2013) Diffusion-weighted magnetic resonance imaging detection of basal forebrain cholinergic degeneration in a mouse model. Neuroimage 66C:133-141. doi: 10.1016/j.neuroimage.2012.10.075
Summary: The authors examined the effectiveness of diffusion MRI using diffusion tensor imaging (DTI) and probabilistic tractography in detecting cholinergic loss in a mouse model. Mice received bilateral 0.2-μg icv injections of mu p75-SAP (Cat. #IT-16). Rabbit IgG-SAP (Cat. #IT-35) was used as control. The animals were then examined using DTI. The data indicate that DTI is a valid technique for assessment of cholinergic loss in septo-hippocampal tracts as a result of Alzheimer's disease.
Krajewski-Hall SJ, Mittelman-Smith MA, Williams H, Lafrance KJ, Mcmullen NT, Rance NE (2012) A role for kisspeptin/neurokinin B/dynorphin (KNDy) neurons in the regulation of estrous cycles and the estrogen modulation of body temperature. Neuroscience 2012 Abstracts 585.02. Society for Neuroscience, New Orleans, LA.
Summary: We have recently described a method to selectively ablate kisspeptin/neurokinin B/dynorphin (KNDy) neurons using stereotaxic injections of NK3-SAP, a neurokinin 3 receptor agonist conjugated to saporin (Mittelman-Smith, Endocrinology, 2012). These studies revealed a critical role for arcuate KNDy neurons in tonic gonadotropin secretion, the rise in serum LH after ovariectomy and estrogen modulation of body weight. Here we determine the effects of KNDy neuron ablation on estrous cycles and the estradiol modulation of body temperature. In the first study, stereotaxic injections of NK3-SAP or Blank-SAP were made in the arcuate nucleus of ovary-intact, adult female rats. Rats with nearly complete KNDy-neuron ablation (verified by NKB immunohistochemistry) exhibited constant diestrus and ovarian atrophy, confirming the importance of these neurons in reproductive regulation. In a second experiment, we evaluated the effects of KNDy neuron ablation on the thermoregulatory axis in rats that were ovariectomized (OVX) and then treated with 17β-estradiol (E2). Tail skin temperatures (TSKIN) and core temperatures (TCORE) were recorded in rats throughout the light/dark cycle and during exposure to different ambient temperatures (TAMBIENT) in an environmental chamber. Notably, the average TSKIN of KNDy-ablated rats was consistently lower than control rats, indicative of lower levels of cutaneous vasodilatation. Moreover, KNDy neuron ablation blocked the reduction of TSKIN by E2 that occurred during the light phase in the environmental chamber, but did not affect the E2 suppression of TSKIN during the dark phase. At a high TAMBIENT of 33 C, the mean TCORE of OVX control rats increased to 39.0 C, and was reduced by E2 replacement. In contrast, at this high TAMBIENT, the average TCORE of OVX, KNDy-ablated rats was lower than OVX control rats, and TCORE was not altered by E2 replacement. Because KNDy neurons exhibit dramatic changes in morphology and gene expression in postmenopausal women, we have hypothesized these neurons contribute to the generation of hot flushes. These studies support this hypothesis by providing the first evidence that KNDy neurons participate in the E2 modulation of body temperature and promote cutaneous vasodilatation, one of the cardinal signs of a hot flush.
Kato J, Satake N, O’Donnell RT, Abuhay M, Lewis C, Tuscano JM (2013) Efficacy of a CD22-targeted antibody-saporin conjugate in a xenograft model of precursor-B cell acute lymphoblastic leukemia. Leuk Res 37(1):83-88. doi: 10.1016/j.leukres.2012.09.010
Summary: Most cases of acute lymphoblastic leukemia (ALL) are of B-cell lineage. Although children with ALL have a high survival rate, there is a subset of children with a much lower survival rate, and long-term side effects from treatment are problematic. CD22 has been suggested as a therapeutic target because it is not present on hematopoietic stem cells, therefore allowing regeneration of normal B cells following depletion of malignant B cells. The authors used a custom conjugate of the antibody HB22.7 and saporin to demonstrate specific toxicity against pre-B ALL cell lines. Mouse IgG-SAP (Cat. #IT-18) was used as a control.
Laplante F, Zhang ZW, Huppe-Gourgues F, Dufresne MM, Vaucher E, Sullivan RM (2012) Cholinergic depletion in nucleus accumbens impairs mesocortical dopamine activation and cognitive function in rats. Neuropharmacology 63(6):1075-1084. doi: 10.1016/j.neuropharm.2012.07.033
Summary: Current thought is that loss of cholinergic function in the nucleus accumbens (N.Acc) is associated with schizophrenia. This deficit is accompanied by low dopaminergic activity in the prefrontal area, which adversely affects working memory. Rats received bilateral injections totaling 500 ng of anti-ChAT-SAP (Cat. #IT-42) into the N.Acc; rabbit IgG-SAP (Cat. #IT-35) was used as a control. Lesioned animals had markedly reduced mesocortical dopamine activation, which corresponded with cognitive impairments. The data suggest that loss of cholinergic neurons in the N.Acc causes loss of dopamine function in the mesocorticolimbic system.
Coolen LM (2012) Featured Article: A pivotal role of lumbar spinothalamic cells in regulation of ejaculation via intraspinal connections. Targeting Trends 13(3)
Mishra SK, Holzman S, Hoon MA (2012) A nociceptive signaling role for neuromedin B. J Neurosci 32(25):8686-8695. doi: 10.1523/JNEUROSCI.1533-12.2012
Summary: Previous work suggests that neuromedin B (NMB) is involved in nociception. Direct injection of the peptide causes nociceptive sensitization, while NMB antagonists attenuate sensitization in reponse to nerve stimulation with mustard oil. Specific subsets of dorsal horn interneurons were eliminated by administering either 10 μg of the custom conjugate neuromedin B-SAP, 0.13 μg of SSP-SAP (Cat. #IT-11), or 1.3 μg of bombesin-SAP (Cat. #IT-40). Blank-SAP (Cat. #IT-21) was used as a control. The data indicate that NMB may be involved in the perception of thermal sensation, but not mechanical or pruritic sensation.
Li AJ, Wiater MF, Oostrom MT, Smith BR, Wang Q, Dinh TT, Roberts BL, Jansen HT, Ritter S (2012) Leptin-sensitive neurons in the arcuate nuclei contribute to endogenous feeding rhythms. Am J Physiol Regul Integr Comp Physiol 302(11):R1313-26. doi: 10.1152/ajpregu.00086.2012
Summary: It is clear that the arcuate nucleus (Arc) plays an important role in the generation of feeding rhythms. To clarify how this region modulates signals governing food intake the authors took advantage of the Arc mediation of leptin. Rats received bilateral injections of leptin-SAP (Cat. #IT-47, 56.5 ng per dose) into each Arc. Blank-SAP (Cat. #IT-21) was used as a control. The lesioned animals quickly became obese and displayed arrhythmic eating patterns under normal light conditions. The results indicate that lesioned neurons in the Arc as well as those in the suprachiasmatic nuclei are required for maintenance of feeding rhythms controlled by photic cues.
Smith CD, Holschbach MA, Olsewicz J, Lonstein JS (2012) Effects of noradrenergic alpha-2 receptor antagonism or noradrenergic lesions in the ventral bed nucleus of the stria terminalis and medial preoptic area on maternal care in female rats. Psychopharmacology (Berl) 224(2):263-276. doi: 10.1007/s00213-012-2749-2
Summary: The authors investigated the function of norepinephrine in mothering. Lesioned animals received 55-ng infusions of anti-DBH-SAP (Cat. #IT-03) into the ventral bed nucleus of the stria terminalis. Mouse-IgG-SAP (Cat. #IT-18) was used as a control. The results demonstrate that downregulated noradrenergic activity is necessary for postpartum maternal behavior, but is not enough to elicit maternal behavior in nulliparous females.
Man SH, Geranton SM, Hunt SP (2012) Lamina I NK1 expressing projection neurones are functional in early postnatal rats and contribute to the setting up of adult mechanical sensory thresholds. Mol Pain 8(1):35. doi: 10.1186/1744-8069-8-35
Summary: Projections from lamina I neurons regulate mechanical and thermal sensitivity due to injury. Little is known about how these pathways develop immediately after birth. Rats at postnatal day 3 were treated with 2 μl of 5 μM SP-SAP (Cat. #IT-07) injected into the intrathecal space. Blank-SAP (Cat. #IT-21) was used as a control. The data show that neurokinin-1 positive neurons project to the parabrachial nucleus in the hindbrain, and that these neurons and lamina I neurons were responsive to noxious stimulation at postnatal day 3. Treated animals also displayed increased mechanical sensitivity from postnatal day 45 on.
Mittelman-Smith MA, Williams H, Krajewski-Hall SJ, Lai J, Ciofi P, McMullen NT, Rance NE ( 2012 ) Arcuate kisspeptin/neurokinin b/dynorphin (KNDy) neurons mediate the estrogen suppression of gonadotropin secretion and body weight. Endocrinology 153(6):2800-2812 . doi: 10.1210/en.2012-1045
Featured Article: Use of a novel saporin conjugate (NK3-SAP) to study the function of neurokinin 3 receptor (NK3r)-expressing kisspeptin/neurokinin B/dynorphin (KNDy) neurons in the rat arcuate nucleus
Rance NE, Mittelman-Smith MA, Krajewski-Hall SJ (2012) Featured Article: Use of a novel saporin conjugate (NK3-SAP) to study the function of neurokinin 3 receptor (NK3r)-expressing kisspeptin/neurokinin B/dynorphin (KNDy) neurons in the rat arcuate nucleus. Targeting Trends 13(2)
Cai L, Gibbs RB, Johnson DA (2012) Recognition of novel objects and their location in rats with selective cholinergic lesion of the medial septum. Neurosci Lett 506(2):261-265. doi: 10.1016/j.neulet.2011.11.019
Summary: This work examined object recognition and object location recognition as specific components of memory. Rats received 0.22 μg of 192-IgG-SAP (Cat. #IT-01) infused into the medial septum followed by testing in novel object recognition (NOR) and object location recognition (OLR) models. Substantial decreases in choline acetyltransferase activity in the hippocampus and frontal cortex produced no difference in NOR but caused a significant impairment in OLR – highlighting the role that septo-hippocampal cholinergic projections play in OLR.
Tolner EA, Sheikh A, Yukin AY, Kaila K, Kanold PO (2012) Subplate neurons promote spindle bursts and thalamocortical patterning in the neonatal rat somatosensory cortex. J Neurosci 32(2):692-702. doi: 10.1523/JNEUROSCI.1538-11.2012
Summary: Immature cortices in both human and rat have spontaneous activity associated with the maturation of cortical synapses and neuronal circuits. In order to investigate what cells are controlling these events the authors administered 400 ng of 192-IgG-SAP (Cat. #IT-01) to the S1 cortex hindlimb/forelimb area of rats. mu p75-SAP (Cat. #IT-16) and mouse-IgG-SAP (Cat. #IT-18) were used as controls. This lesion eliminates subplate neurons which results in a significant loss of evoked spindle burst activity.
St Peters M, Sarter M (2012) Featured Article: Motivation’s modulation of attention through the mesolimbic-corticopetal cholinergic circuitry. Targeting Trends 13(1)
Khan AM KKL, Sanchez-Watts, Ponzio TA, Kuzmiski JB, Bains JS, Watts AG (2011) MAP kinases couple hindbrain-derived catecholamine signals to hypothalamic adrenocortical control mechanisms during glycemia-related challenges. J Neurosci 31(50):18479-18491. doi: 10.1523/JNEUROSCI.4785-11.2011
Summary: This work uses in vivo and ex vivo techniques to clarify how hypothalamic afferent pathways use intracellular mechanisms to modulate glycemia related adrenocortical responses. Rats received 42 ng injections of anti-DBH-SAP (Cat. #IT-03) into the paraventricular nucleus of the hypothalamus. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The results establish a relationship between neurons from nutrient-sensing regions and intracellular mechanisms in hypothalamic corticotropin-releasing hormone neuroendocrine neurons.
Janiesch PC, Kruger HS, Poschel B, Hanganu-Opatz IL (2011) Cholinergic control in developing prefrontal-hippocampal networks. J Neurosci 31(49):17955-17970. doi: 10.1523/JNEUROSCI.2644-11.2011
Summary: In this work the authors examined the role of acetylcholine in the maturation of cognitive processing due to oscillatory rhythms entraining neuronal networks. Rats received 50 ng of 192-IgG-SAP (Cat. #IT-01) into each lateral ventricle, or 25 ng directly into the medial septum. Among other results, cholinergic input was shown to reach the prefrontal cortex toward the end of the first postnatal week, initially targeting GABAergic neurons. Reduction of this activity by lesioning cholinergic neurons may cause global diminishment of neocortical activity.
Botly LC, De Rosa E (2012) Impaired visual search in rats reveals cholinergic contributions to feature binding in visuospatial attention. Cereb Cortex 22(10):2441-2453. doi: 10.1093/cercor/bhr331
Summary: Previous work established the role of acetylcholine from the nucleus basalis magnocellularis in attentional processing and visuospatial attention. In order to investigate the necessity of cortical cholinergic input for support of feature binding in visuospatial attention the authors administered bilateral intraparenchymal injections of 192-IgG-SAP (Cat. #IT-01, 4 injections, 40 ng per injection). Lesioned animals took longer to locate targets during type-specific search trials, demonstrating that cholinergic input influences feature binding during visuospatial attention tasks.
Kozicz T, Xu L, Geenen B, Gaszner B, Kovacs K, Roubos E (2011) Leptin-receptor-expressing neurons in the non-preganglionic Edinger-Westphal nucleus regulate white and brown adipose tissue. Neuroscience 2011 Abstracts 822.19. Society for Neuroscience, Washington, DC.
Summary: Leptin, produced by white adipose tissue (WAT), is a key factor that regulates food intake and energy expenditure in vertebrates. It conveys information about fat storage in the periphery to the brain. The leptin receptor long form (LepRb) can be found in the non-preganglionic Edinger-Westphal nucleus (npEW) in the midbrain, which is the main site of urocortin 1 (Ucn1) production in the brain. In both mice and rats, intraperitoneal administration of leptin induces an increase in Ucn1 expression in the npEW whereas in mice that lack LepRb (db/db mice), the npEW contains considerably reduced amount of Ucn1. The npEW also responds to acute thermal exposure, indicating a role of this nucleus in thermoregulation. Brown adipose tissue (BAT) is critical to maintain homoeothermia and is centrally controlled via sympathetic outputs. A recent study demonstrates a projection from EW to BAT by using retrograde tracer pseudorabies virus (PRV). In our study, using PRV injection into the WAT of rats, we identified PRV-labeled Ucn1 neurons in the npEW, indicating a connection from npEW to WAT. In order to analyze the involvement of the npEW in the regulation of sympathetic WAT and BAT outputs, we performed the experiment using the neurotoxin saporin. When conjugated to leptin (Lep-SAP), Lep-SAP can selectively kill LepRb-expressing neurons. Wister rats were given injection of either Lep-SAP or a control blank saporin (B-SAP) into the left npEW Results showed that injection of Lep-SAP significantly blunted Ucn1 expression in the npEW. The weights of WAT and BAT were analyzed on both sides. The WAT and BAT weights were increased significantly on the contralateral side in Lep-SAP compared with B-SAP injected rats, however not different on the ipsilateral side. Interestingly, we observed that both WAT and BAT weighed more on the ipsilateral than the contralateral side only in the B-SAP animals. We will further test the effect of lesioning npEW neurons on the function of WAT and BAT by assessing specific WAT and BAT markers by RT-PCR and histology. Taken these data together, we provide evidence that LepRb-expressing neurons in the npEW regulate BAT and WAT most probably via sympathetic circuits.
Krajewski SJ, Smith MA, Williams H, Ciofi P, Lai JY, Mcmullen NT, Rance NE (2011) Ablation of NK3 receptor-expressing KNDy neurons in the rat arcuate nucleus using [MePhe7]Neurokinin B-Saporin. Neuroscience 2011 Abstracts 712.09. Society for Neuroscience, Washington, DC.
Summary: A subpopulation of neurons expressing kisspeptin, neurokinin B and dynorphin (KNDy neurons) has been shown to reside within the arcuate nucleus of many mammalian species. Although these peptides are critical for reproductive function, the precise role of the arcuate KNDy neurons is not fully understood. Here we describe a method to ablate KNDy neurons based on their co-expression of the Neurokinin 3 receptor (NK3R, Burke et al., J. Comp. Neurol, 2006). Saporin, a molecular neurotoxin, was conjugated to [MePhe7]Neurokinin B, a selective NK3R agonist ([MePhe7]NKB-SAP, Advanced Targeting Systems, San Diego, CA). Binding studies revealed that the conjugation of saporin did not alter the affinity of [MePhe7]NKB to NK3R in rat cerebral cortex membranes. To investigate the specificity of this conjugate for ablation of NK3R neurons, stereotaxic surgery was used to bilaterally inject [MePhe7]NKB-SAP into the arcuate nucleus of female rats. Control rats were injected with saporin conjugated to a scrambled peptide (Blank-SAP, Advanced Targeting Systems). Rats were sacrificed 31-34 days later and the brains were processed for immunohistochemical studies. Nissl stained sections from [MePhe7]NKB-SAP-treated rats showed no signs of inflammation at the injection sites and no qualitative changes in cell density compared to Blank-SAP control rats. Immunohistochemistry revealed near-complete loss of NK3R-immunoreactive (ir) neurons throughout the arcuate nucleus of [MePhe7]NKB-SAP rats. When the injection site was dorsal to the arcuate nucleus, there was also variable loss of NK3R-ir cells in the lateral hypothalamus and zona incerta. In the arcuate nucleus, [MePhe7]NKB-SAP injections resulted in a 98% and 94% reduction in the number of kisspeptin and neurokinin B-ir neurons, respectively, compared to Blank-SAP controls. The number of dynorphin-ir neurons in the arcuate nucleus of [MePhe7]NKB-SAP-treated rats was reduced by 67%, a value consistent with the co-expression of NK3R on dynorphin neurons in our previous study (Burke et al., J. Comp. Neurol, 2006). In contrast, arcuate proopiomelanocortin and neuropeptide Y immunoreactivity were preserved in [MePhe7]NKB-SAP rats. Moreover, there was no difference in GnRH-ir fiber density in the median eminence between the two groups. These results document the utility of [MePhe7]NKB-SAP for selective ablation of NK3R-expressing KNDy neurons in rat hypothalamus. These rats were used to examine the role of KNDy neurons in the estrogen regulation of LH secretion and body weight in the female rat (see Smith et al., Soc. Neurosci. Abstr. 2011).
Smith MA, Williams H, Krajewski SJ, Mcmullen NT, Rance NE (2011) Arcuate NK3 receptor-expressing KNDy neurons are essential for estrogen modulation of LH secretion and body weight in the female rat. Neuroscience 2011 Abstracts 712.07. Society for Neuroscience, Washington, DC.
Summary: Arcuate kisspeptin, neurokinin B, and dynorphin (KNDy) neurons have been proposed to mediate estrogen negative feedback in multiple species. To determine if these neurons are essential for this feedback, we ablated KNDy neurons in the arcuate nucleus of female rats using [MePhe7]Neurokinin B, a selective NK3 receptor (NK3R) agonist, conjugated to Saporin ([MePhe7]NKB-SAP, Advanced Targeting Systems, San Diego, CA). The specificity of this conjugate for NK3R-expressing KNDy neurons is described in a separate abstract (see Krajewski et al., Soc. Neurosci. Abstr. 2011). Twenty-four female rats were ovariectomized (OVX) and received bilateral arcuate microinjections of either [MePhe7]NKB-SAP or a scrambled peptide conjugated to Saporin (Blank-SAP controls). 20-23 days later, animals were implanted with s.c. silastic capsules containing 17β-estradiol (E2), and animals were sacrificed 11 days later. Blood samples for RIA of serum LH were taken at time of OVX and injections (baseline), 20-23 days post-OVX, and 11 days after E2-treatment. Because OVX and E2-treatment have well-described effects on body weight, animals were weighed at the same three time points. In control animals, OVX induced a 13-fold rise in serum LH, which returned to baseline 11 days after E2 replacement. In contrast, OVX had no effect on serum LH in [MePhe7]NKB-SAP animals. There was a small decrease in serum LH 11 days after E2 replacement in [MePhe7]NKB-SAP animals, but the magnitude of this change was much less than seen in control animals. Control animals also exhibited a 20% increase in body weight 20-23 days after OVX, followed by a significant reduction after E2 replacement. Surprisingly, neither OVX nor E2 replacement affected body weight in [MePhe7]NKB-SAP-treated animals. Rather, these animals showed a steady increase in body weight throughout the experiment, at rates comparable to intact female rats or OVX rats treated with E2 (Williams et al., Endocrinology, 2010). Immunohistochemical studies showed near-complete destruction of KNDy neurons in the arcuate nucleus of [MePhe7]NKB-SAP animals. There was preservation of proopiomelanocortin and neuropeptide Y immunoreactivity in the arcuate nucleus and GnRH-immunoreactive fibers in the median eminence. These data provide compelling evidence that arcuate KNDy neurons play an essential role in estrogen negative feedback on LH secretion as well as the estrogen modulation of body weight.
Talman WT, Nitschke Dragon D, Jones S, Moore SA, Lin L-H (2011) Cardiovascular dysfunction and cardiac injury result from selective glial damage in the nucleus tractus solitarii. Neuroscience 2011 Abstracts 664.14. Society for Neuroscience, Washington, DC.
Summary: In man, extensive CNS dysfunction as may occur after subarachnoid hemorrhage may lead to cardiac damage and cardiac arrhythmias. We have shown that highly selective and restricted lesions of the nucleus tractus solitarii (NTS) may lead to similar cardiac and cardiovascular compromise. For example, using conjugates including the cytotoxin saporin (SAP) to selectively damage NTS neurons that express NK1 receptors or those that express tyrosine hydroxylase (TH) leads to cardiac dysfunction and associated lability of arterial pressure. In continuing efforts to better characterize cellular changes produced by introducing into the NTS conjugates containing SAP, we have studied the effect of anti-dopamine-beta-hydroxylase (anti-DBH)-SAP, stabilized substance P (SSP)-SAP, SAP (unconjugated), blank-SAP (non-targeted peptide conjugate), IgG-SAP (non-targeted immunoglobulin conjugate), and 6-hydoxydopamine (6-OHDA) as a control without SAP injected into NTS. We assessed effects of the injected agents both on cellular markers [NMDAR1 (NMDA receptor subunit 1), GluR2 (AMPA receptor subunit 2), gamma-aminobutyric acid (GABA) receptor type a and b, neuronal nitirc oxide synthase (nNOS), TH, vesicular glutamate transporters (VGluTs), choline acetyl transferase (ChAT), glial fibrillary acidic protein (GFAP), connexin 43 (Cx43), DBH and protein gene product 9.5 (PGP 9.5)] and on cardiovascular and cardiac function. We have found that each compound containing SAP (including blank-SAP, IgG-SAP, unconjugated SAP) led to loss of GFAP and Cx43 immunofluorescent labeling in the NTS as well as lability of arterial pressure, cardiac arrhythmias, and cardiac myocytolysis. Those outcomes occurred despite neuronal specificity for each of the SAP conjugates. For example, anti-DBH-SAP led to a decrease in TH and DBH staining as well as a profound loss in GFAP and Cx43. In contrast, SSP-SAP led to loss of NK1 as well as GFAP, Cx43, and glutamate receptor markers but did not lead to loss of DBH or GABA. SSP-SAP also caused a loss in PGP9.5 which was not observed in all other agents. SAP and blank-SAP, on the other hand, led to loss of GFAP and Cx43 while 6-OHDA led to loss of TH and DBH, increased GFAP and decreased Cx-43. We are still investigating the effects of 6-OHDA on lability of arterial pressure and cardiac events but preliminary data suggest that, in doses used, it led to loss of TH and DBH but did not lead to either lability or cardiac events that were seen with each of the conjugates containing an SAP moiety. This study suggests that glial dysfunction may alone interefere with cardiovascular control through the NTS and may lead to cardiac damage and cardiovascular dysfunction.
Wiater MF, Jansen H, Oostrom M, Li A-J, Dinh T, Ritter S (2011) Lesions targeting leptin-sensitive neurons in the mediobasal hypothalamus dissociate activity and temperature circadian rhythms. Neuroscience 2011 Abstracts 396.11. Society for Neuroscience, Washington, DC.
Summary: Previously we investigated the role of NPY and leptin sensitive networks in the mediobasal hypothalamus in sleep and feeding and found profound regulatory and circadian deficits. We propose that the MBH, particularly the arcuate nuclei (Arc), is required for the integration of homeostatic circadian systems including temperature and activity. We tested this hypothesis with the use of the saporin toxin conjugated to leptin (Lep-SAP) or a blank molecule with no known biological function or receptor (B-SAP) directed to the Arc. Lep-SAP binds to, is internalized by and destroys leptin receptor expressing neurons at the injection site. Lep-SAP rats became obese and hyperphagic and progressed through a dynamic phase to a static phase of growth similar to a ventromedial lesioned rat. Activity and temperature data were collected using intraperitoneal PDT-4000 Emitters with Vital View Data Acquisition Software (Mini Mitter, Philips Respironics, Bend, OR). Circadian rhythms were examined over 49 days during the static phase of obesity in B-SAP (n=10) and Lep-SAP (n=12) rats. Rats were maintained on a 12:12 light:dark (LD) schedule for 13 days and thereafter maintained in continuous dark (DD). After the first thirteen days of DD, food was restricted to four hours per day from 9AM until 1PM for ten days. Immediately thereafter, rats were fasted for three days to evaluate persistence of food-entrained rhythms. Using ClockLab software (Natick, MA) actograms and tempograms were generated as double raster plots. Lomb-Scargle periodograms were used to assess rhythms and their robustness. We found that Lep-SAP rats were arrhythmic for activity in DD, but that food anticipatory activity was nevertheless entrainable to the restricted feeding schedule and the entrained rhythm persisted during the subsequent 3-day fast. Thus, for activity, the light-entrainable oscillator, but not the food entrainable oscillator, was disabled by the MBH lesion. In contrast, temperature remained rhythmic in DD in the Lep-SAP rats, but did not entrain to restricted feeding. We conclude that the leptin-sensitive network of the Arc and MBH is required for entrainment of activity by photic cues and for entrainment of temperature by food and for the integration of these rhythms.
Liu XY, Liu ZC, Sun YG, Ross M, Kim S, Tsai FF, Li QF, Jeffry J, Kim JY, Loh HH, Chen ZF (2011) Unidirectional cross-activation of GRPR by MOR1D uncouples itch and analgesia induced by opioids. Cell 147(2):447-458. doi: 10.1016/j.cell.2011.08.043
Summary: Recent work has begun to define the different pathways used by itch and pain. This study was designed to investigate whether opioids cause the itch sensation by gastrin releasing peptide receptor activation. Mice received intrathecal injections of bombesin-SAP (Cat. #IT-40) in order to investigate the coexpression of various signaling molecules in the spinal cord. Blank-SAP (Cat. #IT-21) was used as a control. The data suggest that opioid-induced itch is independent of opioid analgesia, and is controlled through a mu-opioid receptor isoform.
King T, Porreca F (2011) Featured Article: Contribution of afferent pathways to nerve injury-induced spontaneous pain and evoked hypersensitivity. Targeting Trends 12(4)
Wiater MF, Mukherjee S, Li AJ, Dinh TT, Rooney EM, Simasko SM, Ritter S (2011) Circadian integration of sleep-wake and feeding requires NPY receptor-expressing neurons in the mediobasal hypothalamus. Am J Physiol Regul Integr Comp Physiol 301(5):R1569-R1583. doi: 10.1152/ajpregu.00168.2011
Summary: Feeding and sleep/wake states interact rhythmically across the circadian cycle. It is suspected that the mediobasal hypothalamic area (MBH) is the site where these rhythms are integrated. The authors administered bilateral 24-ng injections of NPY-SAP (Cat. #IT-28) into the arcuate nucleus in order to eliminate NPY receptor-expressing neurons in the MBH of rats. Blank-SAP (Cat. #IT-21) was used as a control. The results indicate that these neurons are required for the interaction of feeding and sleep/wake timing.
Lemons LL, Wiley RG (2011) Galanin receptor-expressing dorsal horn neurons: role in nociception. Neuropeptides 45(6):377-383. doi: 10.1016/j.npep.2011.08.002
Summary: This work examines the mociceptive role of galanin receptor-1-expressing neurons found in the superficial dorsal horn. 500 ng of galanin-SAP (Cat. #IT-34) was injected into the lumbar intrathecal space of rats; blank-SAP (Cat. #IT-21) was used as a control. The rats were then tested in a series of thermal nociception models. Lesioned animals were less sensitive to heat, suggesting that loss of the gal1r-expressing excitatory interneurons disrupted the pain transmission pathway.
Lin Y, Sarfraz Y, Jensen A, Dunn AJ, Stone EA (2011) Participation of brainstem monoaminergic nuclei in behavioral depression. Pharmacol Biochem Behav 100(2):330-9. doi: 10.1016/j.pbb.2011.08.021
Summary: While the classical model states reductions in central noradrenergic activity produces depression, more recent work has indicated that higher activity in this brain region directly correlates with depression. Using a dopamine-ß-hydroxlase targeted toxin to lesion the locus coeruleus of mice, along with goat-IgG-SAP (Cat. #IT-19) as a control, the authors found that treated animals showed increased resistance to depressive behavior in several tests. The results suggest that monoaminergic lesions are greatly affected by mouse strain, lesion size, and involvement of other neuronal systems.
Related Products: Goat IgG-SAP (Cat. #IT-19)
Burke PG, Neale J, Korim WS, McMullan S, Goodchild AK (2011) Patterning of somatosympathetic reflexes reveals nonuniform organization of presympathetic drive from C1 and non-C1 RVLM neurons. Am J Physiol Regul Integr Comp Physiol 301(4):R1112-R1122. doi: 10.1152/ajpregu.00131.2011
Summary: Some neurons in the rostral ventrolateral medulla are part of the circuitry that helps maintain blood pressure. This control is exerted through both feed-forward and reflex adjustment mechanisms. The authors used bilateral injections of anti-DBH-SAP (Cat. #IT-03, 24 ng per side) into the spinal cord of rats between T1 and T2 to better understand the organization of this circuitry. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The results suggest that myelinated neurons may control baseline tone, while stressor response uses unmyelinated neurons.
Radley JJ, Sawchenko PE (2011) A common substrate for prefrontal and hippocampal inhibition of the neuroendocrine stress response. J Neurosci 31(26):9683-95. doi: 10.1523/JNEUROSCI.6040-10.2011
Summary: In order to better understand how response to emotional stress is regulated, the authors injected 114 ng of GAT-1-SAP (Cat. #IT-32) into each side of the anterior bed nucleus of the stria terminalis. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The results suggest that medial prefrontal cortex and hippocampal formation influences on stress regulation use the same access to modulate emotional stress rather than having parallel networks.
St Peters M, Demeter E, Lustig C, Bruno JP, Sarter M (2011) Enhanced control of attention by stimulating mesolimbic-corticopetal cholinergic circuitry. J Neurosci 31(26):9760-9771. doi: 10.1523/JNEUROSCI.1902-11.2011
Summary: Motivation and attention interact to preserve cognitive performance under challenging conditions. In order to better define the circuitry connecting these two processes, the authors lesioned the prefrontal cortex (200 ng of 192-IgG-SAP, Cat. #IT-01) and the posterior parietal cortex (280 ng of 192-IgG-SAP). Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data indicate that cholinergic projections to the cortex modulate detection of clues and filtering of distractors during attentional tasks, accentuating cognitive control.
King T, Qu C, Okun A, Mercado R, Ren J, Brion T, Lai J, Porreca F (2011) Contribution of afferent pathways to nerve injury-induced spontaneous pain and evoked hypersensitivity. Pain 152(9):1997-2005. doi: 10.1016/j.pain.2011.04.020
Summary: Whether exaggerated pain response to a normally innocuous tactile stimulus should be defined as allodynia has been debated. Through the use of several techniques, one of which was intrathecal injection of SSP-SAP (Cat. #IT-11, 16.5 pg), the authors examined which pathways were utilized in this type of pain. Blank-SAP (Cat. #IT-21) was used as a control. The data indicate that tactile stimulation may reflect a different pain state than allodynia.
Wilkinson KA, Fu Z, Powell FL (2011) Ventilatory effects of substance P-saporin lesions in the nucleus tractus solitarii of chronically hypoxic rats. Am J Physiol Regul Integr Comp Physiol 301(2):R343-R350. doi: 10.1152/ajpregu.00375.2010
Summary: Interaction of the multiple brainstem areas that have been established as CO2-sensitive is not well understood. In order to investigate chemoreceptor roles in the nucleus tractus solitarii (NTS) the authors injected 2.6 ng of SP-SAP (alternative: SSP-SAP; Cat. #IT-11) into the caudal NTS of rats. Blank-SAP (Cat. #IT-21) was used as a control. The results indicate that neurokinin-1 receptor-expressing cells in the NTS contribute to plasticity during chronic hypoxia.
Laplante F, Lappi DA, Sullivan RM (2011) Cholinergic depletion in the nucleus accumbens: Effects on amphetamine response and sensorimotor gating. Prog Neuropsychopharmacol Biol Psychiatry 35(2):501-509. doi: 10.1016/j.pnpbp.2010.12.005
Summary: Disruption of dopamine and acetylcholine balance in the striatum may play a role in conditions such as Parkinson's and schizophrenia. In this work the authors lesioned cholinergic neurons in the nucleus accumbens (N.Acc) with the novel toxin Anti-ChAT-SAP (Cat. #IT-42). Rats received 0.25-µg bilateral injections of the toxin into the N.Acc. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The results of this lesion produced responses that may parallel the loss of cholinergic neurons seen in schizophrenia.
Goehler LE, Gaykema RPA (2011) Featured Article: Targeted lesion of caudal brainstem catecholamine neurons reveals their role in symptoms of fatigue. Targeting Trends 12(1)
Zhang L, Hadley GA (2010) Application of anti-CD103 immunotoxin for saving islet allograft in context of transplantation. Chin Med J (Engl) 123(24):3644-51.
Summary: This work investigates whether depletion of CD103-positive cells protects transplanted islets from host-immune cell attack. Diabetes was induced in mice, followed by an islet transplant. Anti-CD103-SAP (Cat. #IT-50) was administered via i.p. injection (1.0 mg/kg or 2.0 mg/kg). Rat IgG-SAP (Cat. #IT-17) was used as a control. Diabetic mice treated with anti-CD103-SAP after islet transplantation had an indefinite survival time as compared to untreated mice that survived fewer than 20 days.
Wiater MF, Oostrom M, Barfield R, Dinh TT, Li A-J, Ritter S (2010) Leptin-saporin lesions of the arcuate nucleus induce an arrhythmic circadian feeding pattern. Neuroscience 2010 Abstracts 733.6. Society for Neuroscience, San Diego, CA.
Summary: The endogenous circadian rhythm of feeding is incompletely understood. The leptin sensitive network within the arcuate nucleus (Arc) of the hypothalamus is important for the control of feeding. Genetic deletion of leptin or leptin receptors results in profound obesity, hyperphagia, and the loss of day/night differences in food intake. Because the Arc is critically involved in control of food intake and contains leptin receptors, we hypothesized that the Arc plays an important role in maintenance of feeding rhythms. To examine this hypothesis, we injected a newly developed targeted toxin, leptin conjugated to saporin (LSAP), into the Arc to lesion leptin receptor-expressing neurons in the vicinity of the injection. Controls were injected saporin conjugated to a peptide with no known action or receptor (blank-saporin, BSAP). We expected the Arc LSAP would disrupt the circadian rhythm of food intake, as seen in rats with genetic deletion of leptin or its receptor. Eating rhythms were monitored continuously (each minute) over a 60-day period using BioDAQ (Research Diets) automated meal monitoring equipment. Data were analyzed for circadian rhythm using ClockLab (ActiMetrics) software. Eatograms (food intake in actogram format), showing eating times and durations comparable to actograms used for wheel-running activity, and Chi-square periodograms were generated. Feeding was monitored in light:dark, dark:dark, or light:light conditions. The LSAP injection caused profound hyperphagia, weight gain and disrupted circadian feeding patterns. Although LSAP rats remained sensitive to light and dark under certain circumstances and were capable of an apparent rhythm during light:dark conditions, feeding was arrhythmic by all measures when photic cues were removed (i.e., in dark:dark and light:light conditions). At the end of experimentation, lesions were analyzed using immunohistochemistry to detect agouti gene related protein (AGRP) and α-melanocortin stimulating hormone (α-MSH) neurons, both known to express leptin receptors. Cell bodies positive for these peptides were greatly diminished in the Arc. Results indicate that the Arc contributes importantly to the expression of circadian rhythms of food intake.
Wiater MF, Mukherjee S, Dinh TT, Rooney E, Li A-J, Simasko SM, Ritter S (2010) The arcuate nucleus of the hypothalamus controls the circadian distribution of sleep and feeding. Neuroscience 2010 Abstracts 648.16/H17. Society for Neuroscience, San Diego, CA.
Summary: Integration of daily sleep and feeding rhythms is incompletely understood. We examined the role of the hypothalamic arcuate nucleus (Arc) in these processes using Arc microinjections of the targeted toxin, NPY-saporin (NPY-sap), or control blank-saporin (B-sap). NPY-sap targets and destroys NPY receptor-expressing neurons. We monitored 24 hr feeding over a 30-day period beginning 2 wks after the Arc injections, and used EEG recordings to assign vigilance states. Vigilance was divided into rapid-eye movement sleep (REMS), non-REMS (NREMS) and wake. NPY-sap lesioned rats were hyperphagic , consuming up to 225% of pre-injection baseline. They rapidly became obese. While in the sleep-monitoring chambers, their body weight change per week ranged from 56 ± 9 g to 40.5 ± 4.5g, compared to 6 ± 0.4 g/wk for B-sap rats. Their circadian pattern of food intake was severely disrupted, such that intake in light and dark periods were approximately equal (43% of their total intake was consumed in the light period vs. 25% in B-sap controls). Sleep patterns were also significantly disrupted in the NPY-sap animals. The occurrence of rapid eye movement sleep (REMS) was inverted in phase, occurring mainly at night, rather than during the day. NonREMS was distributed equally across day and night, instead of occurring predominantly during the day. However, 24-hr total REMS and NREMS time was normal. B-sap controls had normal sleep patterns, with NREMS and REMS occurring predominantly in the light phase. To determine if the change in sleep pattern was due to the change in feeding patterns, we restricted access to food to the dark period for 4 days. NPY-sap treated animals doubled their food intake in the dark period. However, sleep patterns were not changed compared to the ad libitum feeding period in either NPY-sap or B-sap rats. After 7 days of ad libitum feeding, we restricted food access to the light period for 4 days. Again, NPY-sap animals doubled their intake during the feeding period, this time during the light phase, and sleep patterns were not changed in either group by the restricted feeding. By 100 days post-lesion, the NPY-sap animals were still obese, but the patterning and amount of their food intake were becoming similar to controls. However, when evaluated again, sleep patterns were still altered to the same degree as observed early post-lesion. These results confirm the importance of NPY-receptive Arc neurons in controlling food intake. They also reveal an unexpected role for the Arc in the timing of both NREMS and REMS that appears to be independent of the patterning of food intake.
Burke PG, Neale J, Korim WS, Mcmullan S, Pilowsky PM, Goodchild AK (2010) Patterning of somatosympathetic reflexes: Identification of distinct bulbospinal sympathoexcitatory RVLM projections by conduction velocity and catecholamine phenotype. Neuroscience 2010 Abstracts 694.11/HHH34. Society for Neuroscience, San Diego, CA.
Summary: The aim of this study was to examine the somatosympathetic reflex (SSR) response of different sympathetic nerves to identify distinct projections of presympathetic vasomotor RVLM neurons by axonal conduction and catecholamine phenotype. All experiments were conducted in urethane-anaesthetised (1.3 g/kg ip), paralysed, vagotomised and artificially ventilated Sprague Dawley rats (n = 44). First, we determined the simultaneous activity of dorsal root potentials and the splanchnic SSR to single shock sciatic nerve (SN) stimulation (single 0.2 ms pulse, 50 sweeps at 0.5-1 Hz, 1-80 V, n=4). Second, we simultaneously recorded the sympathoexcitatory response of multiple, sympathetic nerves (cervical, renal, splanchnic and lumbar) to low (A-fibre afferent; 4-10 V) and high (A- and C-fibre afferents; +40 V) SN stimulation (n=19). Third, we examined the cervical or splanchnic SSR to low intensity SN stimulation in rats following RVLM microinjection of somatostatin (SST) or muscimol (n=8). Fourth, we examined the splanchnic SSR in rats pretreated with intraspinal anti-dopamine-beta-hydroxylase-saporin (anti-DβH-SAP; 24 ng/side, n=8), a neurotoxin that depleted ~70% of catecholamine (C1) neurons in the RVLM compared to IgG-saporin control (n=5). Low intensity SN stimulation evoked biphasic responses in the renal, splanchnic and lumbar nerves but a single peak in the cervical nerve. High intensity SN stimulation evoked triphasic responses in the renal, splanchnic and lumbar nerves and a biphasic cervical response. RVLM injections of SST abolished the early peak of the cervical and splanchnic SSR. Intraspinal pretreatment with anti-DβH-SAP eliminated the late peak of the splanchnic SSR and attenuated the first peak. It is concluded that the mono- or bi-phasic SSR responses are generated by A-fibre afferent inputs driving two classes of bulbospinal sympathoexcitatory RVLM neurons with myelinated or unmyelinated axonal conduction. Secondly, unmyelinated RVLM presympathetic neurons, presumed to be all C1, innervate splanchnic, renal and lumbar SPN, whereas myelinated C1 and non-C1 neurons innervate all sympathetic outflow examined. These findings extend prior evidence that the RVLM expresses several types of phenotypically distinct descending sympathoexcitatory pathways.
Lyons AM, Thiele TE (2010) Neuropeptide Y conjugated to saporin alters anxiety-like behavior when injected into the central nucleus of the amygdala or basomedial hypothalamus in BALB/cJ mice. Peptides 31(12):2193-2199. doi: 10.1016/j.peptides.2010.09.009
Summary: Neuropeptide Y (NPY) in the hypothalamus is known to modulate feeding behavior. In this work the authors used bilateral 48 ng injections of NPY-SAP (Cat. #IT-28) into the central amygdala or basomedial hypothalamus (BMH) of rats to investigate the role of NPY in anxiety. Blank-SAP (Cat. #IT-21) was used as a control. Injections into the amygdala increased anxiety-like behavior, while injections into the BMH reduced anxiety-like behavior. BMH injections also initiated an increase of NPY-1 receptor expression in the basolateral nuclei of the amygdala.
Di Sebastiano AR, Wilson-Perez HE, Lehman MN, Coolen LM (2011) Lesions of orexin neurons block conditioned place preference for sexual behavior in male rats. Horm Behav 59(1):1-8. doi: 10.1016/j.yhbeh.2010.09.006
Summary: The neuropeptide orexin is important in the feedback mechanisms of food intake and drugs of abuse. This work investigates the role of orexin in sexual reward in male rats. Two 200 ng bilateral hypothalamic injections of orexin-SAP (Cat. #IT-20) were made into each hemisphere. Blank-SAP (Cat. #IT-21) was used as a control. Although it was shown orexin neurons are activated by sexual reward cures, the data suggest that orexin is not essential for sexual performance and motivation.
Baskin DG, Kim F, Gelling RW, Russell BJ, Schwartz MW, Morton GJ, Simhan HN, Moralejo DH, Blevins JE (2010) A new oxytocin-saporin cytotoxin for lesioning oxytocin-receptive neurons in the rat hindbrain. Endocrinology 151(9):4207-4213. doi: 10.1210/en.2010-0295
Summary: There is evidence to suggest that release of oxytocin in the nucleus tractus solitarius (NTS) of the hindbrain can inhibit food intake by augmenting the cholecystokinin satiety response. In order to add support to this hypothesis the authors used oxytocin-SAP (Cat. #IT-46) to eliminate oxytocin receptive cells in the NTS. Blank-SAP (Cat. #IT-21) was used as a control. 0.5 µl-injections of oxytocin-SAP into the NTS caused reduced satiation effect of CCK-8 and blocked the stimulation of food intake by an oxytocin receptor antagonist.
Northrop LE, Polston EK, Erskine MS (2010) Noradrenergic nuclei that receive sensory input during mating and project to the ventromedial hypothalamus play a role in mating-induced pseudopregnancy in the female rat. J Neuroendocrinol 22(10):1061-1071. doi: 10.1111/j.1365-2826.2010.02049.x
Summary: Maintenance of pregnancy or pseudopregnancy in rats is maintained by bicircadian prolactin surges induced by vaginal-cervical stimulation. In order to test the hypothesis that medullary noradrenergic cell groups are involved in this process the authors infused rats with either 2 ng or 60 ng anti-DBH-SAP (Cat. #IT-03) into the ventrolateral division of the ventromedial hypothalamus (VMHv1) and the posterodorsal medial amygdala. Mouse IgG-Sap (Cat. #IT-18) was used as a control. The data confirm that noradrenergic neurons are involved in the maintenance of pregnancy or pseudopregnancy.
de Carvalho D, Bicego KC, de Castro OW, da Silva GS, Garcia-Cairasco N, Gargaglioni LH (2010) Role of neurokinin-1 expressing neurons in the locus coeruleus on ventilatory and cardiovascular responses to hypercapnia. Respir Physiol Neurobiol 172(1-2):24-31. doi: 10.1016/j.resp.2010.04.016
Summary: NK-1 receptors (NK1R) play an important role in cardiorespiratory responses to hypercapnia. In order to paint a clearer picture of the systems involved the authors injected 0.4 µl of 2 µM SP-SAP (Cat. #IT-07) into the locus coeruleus (LC) of rats. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data suggest that several subpopulations of neurons express NK1R in the LC, and that these subpopulations play different roles in the modulation of cardiorespiratory reponses to hypercapnia.
Potes CS, Turek VF, Cole RL, Vu C, Roland BL, Roth JD, Riediger T, Lutz TA (2010) Noradrenergic neurons of the area postrema mediate amylin's hypophagic action. Am J Physiol Regul Integr Comp Physiol 299(2):R623-631. doi: 10.1152/ajpregu.00791.2009
Summary: Amylin decreases food intake in rats and is a satiation signal affecting the area postrema (AP). This work investigated the role of noradrenergic neurons in amylin activity. Rats received a total of 50 ng of anti-DBH-SAP (Cat. #IT-03) into the AP and 25 ng into the lateral parabrachial nucleus. Mouse IgG-SAP (Cat. #IT-18) was used as a control. Rats showing a >50% lesion of noradrenergic neurons were unresponsive to low doses of amylin, suggesting that noradrenergic neurons are part of the amylin pathway.
Di Sebastiano AR, Yong-Yow S, Wagner L, Lehman MN, Coolen LM (2010) Orexin mediates initiation of sexual behavior in sexually naive male rats, but is not critical for sexual performance. Horm Behav 58(3):397-404. doi: 10.1016/j.yhbeh.2010.06.004
Summary: In this work the role of endogenous orexin-A and B in male sexual behavior was investigated. Rats received a total of 400 ng of orexin-SAP (Cat. #IT-20) into the hypothalamus in each hemisphere. Blank-SAP (Cat. #IT-21) was used as a control. The lesions facilitated initiation of sexual behavior in naïve males, and reduced anxiety-like behaviors. The data suggest that orexin may play a role in arousal and anxiety related to sexual behavior in naïve animals, but is not critical for performance or motivation.
Hovinga KE, Shimizu F, Wang R, Panagiotakos G, Van Der Heijden M, Moayedpardazi H, Correia AS, Soulet D, Major T, Menon J, Tabar V (2010) Inhibition of notch signaling in glioblastoma targets cancer stem cells via an endothelial cell intermediate. Stem Cells 28:1019-1029. doi: 10.1002/stem.429
Summary: The antibody CD105 (1:1,000) was incubated with Mab-ZAP, to allow binding and formation of a CD105 antibody-saporin complex, which was added to explants or to a human cerebral microvessel endothelial cell line as control. CD105 antibody was also incubated with a Goat-IgG-SAP for control that does not bind CD105. The conjugates were injected into the explant under a dissecting microscope after gently incising the explant surface for better access.
Vianna DM, Carrive P (2010) Cardiovascular and behavioural responses to conditioned fear and restraint are not affected by retrograde lesions of A5 and C1 bulbospinal neurons. Neuroscience 166:1210-1218. doi: 10.1016/j.neuroscience.2010.01.039
Summary: To investigate the role of A5 neurons in some forms of psychological stress the authors injected 22 or 44 ng of anti-DBH-SAP (Cat. #IT-03) into the spinal cord of rats. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data shows that A5 presympathetic neurons are not essential for the expression of the tachycardic and pressor responses to conditioned fear and restraint.
Dailey MJ, Bartness TJ (2010) Arcuate nucleus destruction does not block food deprivation-induced increases in food foraging and hoarding. Brain Res 1323:94-108. doi: 10.1016/j.brainres.2010.01.078
Summary: While some aspects of food intake are understood, mechanisms controlling hoarding of food have not been identified. This work investigates the role of NPY in the arcuate nucleus (Arc) in hoarding. Siberian hamsters received 48 ng injections of NPY-SAP (Cat. #IT-28) into the Arc; blank-SAP (Cat. #IT-21) was used as a control. In lesioned animals food deprivation-induced hoarding was increased 100%, but baseline foraging and food hoarding was unchanged.
Li AJ, Wang Q, Dinh TT, Ritter S (2010) Featured Article: Deletion of NPY/AGRP and POMC neurons in the arcuate nucleus by leptin-saporin produces hyperphagia, obesity and changes in diurnal feeding patterns in rats. Targeting Trends 11(1)
Chen ZF, Sun YG, Zhao ZQ, Meng XL, Yin J, Liu XY (2009) Featured Article: Ablation of GRPR+ neurons in the spinal cord by bombesin-saporin knocks out itch sensation in mice without affecting pain circuit. Targeting Trends 10(4)
Jinks SL, Carstens E, Antognini JF (2009) Nitrous oxide-induced analgesia does not influence nitrous oxide's immobilizing requirements. Anesth Analg 109:1111-1116. doi: 10.1213/ANE.0b013e3181b5a2a7
Summary: Noradrenergic neurons in the locus coeruleus (LC) are involved with the analgesic action of nitrous oxide (N2O). In order to examine whether these neurons are also involved with the immobilizing effects of N2O, rats received 4 µg intracerebroventricular injections of anti-DBH-SAP (Cat. #IT-03). Mouse IgG-SAP (Cat. #IT-18) was used as a control. Lesioned animals did not experience the analgesic effects of N2O, but the immobilizing effects were still present. The data demonstrate that the immobilizing mechanism of N2O is independent from its analgesic effects.
Dashniani M, Burjanadze M, Beselia G, Maglakelidze G, Naneishvili T (2009) Spatial memory following selective cholinergic lesion of the nucleus basalis magnocellularis. Georgian Med News 174:77-81.
Summary: This study investigated the role of cholinergic nucleus basalis magnocellularis (NBM) cells in learning and memory. Rats received bilateral 200 ng injections of 192 IgG-SAP (Cat. IT-01) into the NBM. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The results indicate that the NBM is important in accurate spatial learning and the processing of information about the spatial environment. Deficits in rats with the cholinergic lesion may be due to lowered attentional function.
Zhang L, Moffatt-Bruce SD, Gaughan AA, Wang JJ, Rajab A, Hadley GA (2009) An anti-CD103 immunotoxin promotes long-term survival of pancreatic islet allografts. Am J Transplant 9:2012-2023. doi: 10.1111/j.1600-6143.2009.02735.x
Summary: The integrin CD103 is suspected of promoting organ allograft rejection and graft-versus-host disease. A custom conjugation was done between the non-depleting aCD103 antibody M290 and saporin. The conjugate was administered at 2.0 mg/kg to mice as an intraperitoneal injection (mouse IgG-SAP, Cat. #IT-18) was used as a control). The mice had previously received an islet transplant into a kidney capsule. Mice treated with M290-SAP were effectively depleted of CD103+ cells and had long term acceptance of the allografts.
Wiley RG, Lemons LL, Kline IV RH (2009) Neuropeptide Y receptor-expressing dorsal horn neurons: role in nocifensive reflex responses to heat and formalin. Neuroscience 161:139-147. doi: 10.1016/j.neuroscience.2008.12.017
Summary: This work examines the effect of lumbar intrathecal administration of NPY-SAP (Cat. #IT-28), and the role of Y1 NPY receptor-expressing neurons (Y1R) in response to thermal and chemical stimulation. Rats received 500 ng or 750 ng intrathecal injections of NPY-SAP. Blank-SAP (Cat. #IT-21) was used as a control. Lesioned animals displayed a specific loss of Y1R in the dorsal horn, as well as reduced nocifensive reflex responses.
Ray AP, Chebolu S, Ramirez J, Darmani NA (2009) Ablation of least shrew central neurokinin NK1 receptors reduces GR73632-induced vomiting. Behav Neurosci 123:701-706. doi: 10.1037/a0015733
Summary: In this work the authors investigated the role of central and peripheral nervous systems components that mediate the emetic reflex. Least shrews received an 600-ng injection of SSP-SAP (Cat. #IT-11) into the lateral ventricle. Some animals also received a 4.8-µg intraperitoneal injection of SSP-SAP. Blank-SAP (Cat. #IT-21) and unconjugated saporin (Cat. #PR-01) were used as controls. Lesioned animals displayed reduced emesis, but the data indicate that a minor peripheral nervous system component is also present.
Radley JJ, Gosselink KL, Sawchenko PE (2009) A discrete GABAergic relay mediates medial prefrontal cortical inhibition of the neuroendocrine stress response. J Neurosci 29:7330-7340. doi: 10.1523/JNEUROSCI.5924-08.2009
Summary: GABAergic neurons have been implicated in the negative regulation of the hypothalamic-pituitary-adrenal axis (HPA). In order to clarify GABAergic input to the paraventricular hypothalamic nucleus the authors injected 0.23 µg of GAT1-SAP (Cat. #IT-32) into the anterior bed nucleus of the stria terminalis. Both unilateral and bilateral injections were used. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data indicate that the GABAergic neuronal population functions as proximate mediator of HPA-inhibitory limbic influences.
Brightwell JJ, Taylor BK (2009) Noradrenergic neurons in the locus coeruleus contribute to neuropathic pain. Neuroscience 160:174-185. doi: 10.1016/j.neuroscience.2009.02.023
Summary: Noradrenergic neurons were eliminated with 5 µg intracerebroventricular injections of anti-DBH-SAP (Cat. #IT-03). Mouse IgG-SAP (Cat. #IT-18) was used as a control. Animals lesioned with anti-DBH-SAP displayed a reduction in behavioral signs of several kinds of allodynia.
Khan AM, Rapp KL, Ponzio TA, Sanchez-Watts G, Watts AG (2009) Featured Article: Deletion of catecholaminergic neurons by anti-DBH-saporin disrupts hypothalamic MAP kinase and CREB activation. Targeting Trends 10(2)
Dashniani MG, Beseliia GV, Maglakelidze GA, Burdzhanadze MA, Chkhikvishvili N (2009) Effects of the selective lesions of cholinergic septohippocampal neurons on different forms of memory and learning process. Georgian Med News 166:81-85.
Summary: The hippocampus is crucial for the ability to recollect everyday events and factual knowledge. Here the authors looked at the role of the septo-hippocampal cholinergic system of the medial septum in learning and memory. Rats received 200 ng injections of 192-IgG-SAP (Cat. #IT-01) into the medial septum. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The data suggest that although the septo-hippocampal region is essential for spatial learning, hippocampal acetyl cholinesterase may not be essential for all types of hippocampal-dependent memory.
Loyd DR, Wang X, Murphy AZ (2008) Sex differences in micro-opioid receptor expression in the rat midbrain periaqueductal gray are essential for eliciting sex differences in morphine analgesia. J Neurosci 28:14007-14017. doi: 10.1523/JNEUROSCI.4123-08.2008
Summary: The authors test whether the periaqueductal gray (PAG), that contains a dense population of µ-opioid receptor (MOR)-expressing neurons, is sexually dimorphic. Rats were injected with 3 pmol of Dermorphin-SAP (Cat. #IT-12) into the PAG. Blank-SAP (Cat. #IT-21) was used as a control. Both behavioral and immunohistochemical evidence suggest that differential expression of MOR-expressing neurons in the PAG between male and female rats accounts for the difference in response to morphine.
Rouleau C, Curiel M, Weber W, Smale R, Kurtzberg L, Mascarello J, Berger C, Wallar G, Bagley R, Honma N, Hasegawa K, Ishida I, Kataoka S, Thurberg BL, Mehraein K, Horten B, Miller G, Teicher BA (2008) Endosialin protein expression and therapeutic target potential in human solid tumors: sarcoma versus carcinoma. Clin Cancer Res 14:7223-7236. doi: 10.1158/1078-0432.CCR-08-0499
Summary: Endosialin is an antigen expressed in many human cancer cell lines. As part of a wide-ranging study investigating clinical specimens, cell culture, and animal models, this group used Hum-ZAP (Cat. #IT-22) combined with a humanized anti-endosialin antibody in cell proliferation assays. Mouse IgG-SAP (Cat. #IT-18) was used as a control. The anti-endosialin antibody and Hum-ZAP were incubated together in equimolar concentrations then applied to cells in culture. Various cancers, including synovial sarcoma, fibrosarcoma, and osteosarcoma among others, were found to express endosialin.
Di Sebastiano AR, Yong-yow S, Coolen LM (2008) The role of orexin in sexual behavior and sexual reward of the male rat. Neuroscience 2008 Abstracts 97.4/UU18. Society for Neuroscience, Washington, DC.
Summary: The hypothalamic neuropeptide orexin has been demonstrated to play a role in reward related to drugs of abuse and is potentially involved in regulation of natural rewarding behaviors. Male sexual behavior has been shown to activate orexin neurons and this behavior is altered by administration of orexin receptor agonists or antagonists. However, the exact role of orexin in male sexual performance, sexual motivation and reward is currently unclear. Therefore, the goal of the current study was to test the hypothesis that orexin plays a critical role in sexual behavior, motivation and reward. First, using Fos as a marker for neural activation, we investigated activation of orexin neurons following different parameters of sexual behavior in sexually naïve and experienced male rats. It was demonstrated that orexin neurons in the lateral hypothalamic area (LHA) and in the dorsal medial hypothalamus/perifornical (PFA-DMH) region become activated with presentation of the female and there is no further increase in activation with other components of mating (15-30% in LHA; 65-80% in PFA-DMH). Next, we tested the functional role of orexin utilizing orexin-cell body specific lesions. Adult male rats underwent lesion or sham surgery using the targeted toxin orexin-saporin or blank-saporin respectively. Following two weeks recovery, sexual behavior was recorded over the course of four mating trials. During the first mating trial, males with complete lesions showed significantly shorter latencies to mount and intromit. This suggests that lesions facilitated sexual performance in naïve animals. This facilitation was attenuated by sexual experience as lesions did not affect any parameter of sexual behavior in experienced animals. Next, runway tests were conducted to determine motivation to run towards a potential partner over two conditioning trials. Lesions did not alter sexual motivation, as lesion and sham males all demonstrated increased speed to run towards an estrous female during the second trial. Finally, a conditioned place preference (CPP) paradigm was conducted as a measure of sexual reward. All groups formed a conditioned preference for the mating-paired chamber, indicating that lesions did not significantly disrupt sexual reward. Overall, these findings suggest that orexin does not play a critical role in male sexual performance, motivation, and reward, however may be involved in general arousal related to sexual behavior.
Likhtik E (2008) Featured Article: Selective lesions of amygdala intercalated neurons using the Dermorphin-SAP immunotoxin reveal their role in conditioned fear. Targeting Trends 9(4)
Göz D, Studholme K, Lappi DA, Rollag MD, Provencio I, Morin LP (2008) Targeted destruction of photosensitive retinal ganglion cells with a saporin conjugate alters the effects of light on mouse circadian rhythms. PLoS ONE 3(9):e3153. doi: 10.1371/journal.pone.0003153
Summary: Retinal ganglion cells expressing melanopsin photopigment are thought to be involved in non-image forming visual responses to light. The authors had a custom conjugate made between saporin and an anti-melanopsin antibody. A 400-ng injection of the melanopsin-SAP conjugate into the eye of a mouse resulted in a 57% loss of the targeted cells. Rabbit IgG-SAP (Cat. #IT-35) was used as a control. The data indicates that melanopsin-containing cells are involved in the response to certain non-image forming visual input.
Likhtik E, Popa D, Apergis-Schoute J, Fidacaro GA, Pare D (2008) Amygdala intercalated neurons are required for expression of fear extinction. Nature 454(7204):642-645. doi: 10.1038/nature07167
Summary: Scientists have been using fear learning in animals to study human anxiety disorders. In order to investigate the contribution of amygdala plasticity to fear learning, rats received 0.25-µl bilateral infusions of 3-µM dermorphin-SAP (Cat. #IT-12) into the amygdala. Blank-SAP (Cat. #IT-21) was used as a control. Lesioned rats displayed extinction expression deficits, indicating that the eliminated intercalated amygdala neurons play a large role in the extinction process.
Radley JJ (2008) Featured Article: Noradrenergic innervation of the dorsal medial prefrontal cortex modulates hypothalamo-pituitary-adrenal responses to acute emotional stress. Targeting Trends 9(3)
Darmani NA, Wang Y, Abad J, Ray AP, Thrush GR, Ramirez J (2008) Utilization of the least shrew as a rapid and selective screening model for the antiemetic potential and brain penetration of substance P and NK1 receptor antagonists. Brain Res 1214:58-72. doi: 10.1016/j.brainres.2008.03.077
Summary: This work investigated the role of central tachykinin NK1 receptors in delayed phase vomiting caused by chemotherapeutics. Least shrews received 1.2 mg/kg intraperitoneal injections of SSP-SAP (Cat. #IT-11). Saporin (Cat. #PR-01) and blank-SAP (Cat. #IT-21) were used as controls. In response to administration of a NK1 receptor agonist lesioned animals vomited less than the control group, indicating an important role for NK1 receptors in emesis.
Li A, Emond L, Nattie E (2008) Brainstem catecholaminergic neurons modulate both respiratory and cardiovascular function. (eds. Poulin MJ, Wilson RJA). In: Integration in Respiratory Control. Advances in Experimental Medicine and Biology 605:371-376. Springer, New York, NY. doi: 10.1007/978-0-387-73693-8_65
Summary: The authors examined the role of brainstem catecholamine (CA) neurons in various aspects of breathing and chemoreception. Rats received 5-µg injections of anti-DBH-SAP (Cat. #IT-03) into the 4th ventricle; mouse IgG-SAP (Cat. #IT-18) was used as a control. This method of lesioning left the CA neurons in the peripheral nervous system intact. Lesioned animals displayed a constant decrease in breathing frequency, reduced response to CO2, and increased variability of breathing during REM sleep. Inhibitory cardiovascular effects were also seen.
Li AJ, Dinh TT, Ritter S (2008) Hyperphagia and obesity produced by arcuate injection of NPY-saporin do not require upregulation of lateral hypothalamic orexigenic peptide genes. Peptides 29(10):1732-1739. doi: 10.1016/j.peptides.2008.05.026
Summary: It has already been shown that lesioning NPY receptor-expressing cells in the arcuate nucleus (Arc) and basomedial hypothalamus produces obesity in rats. The authors examined the contribution of orexigenic peptides, orexins, and melanocortin-concentrating hormone to the lesion effects. Rats received bilateral 24 ng injections of NPY-SAP (Cat. #IT-28) into the dorsal border of the Arc. Blank-SAP (Cat. #IT-21) was used as a control. The data suggest that obesity produced by NPY-SAP lesion is different than dietary obesity or obesity associated with leptin or leptin receptor deficiency.
Young EE, Baumbauer KM, Hillyer JE, Patterson AM, Hoy KC, Jr., Mintz EM, Joynes RL (2008) The neonatal injury-induced spinal learning deficit in adult rats: central mechanisms. Behav Neurosci 122:589-600. doi: 10.1037/0735-7044.122.3.589
Summary: This report examined whether neonatal injuries had any contralateral effects in adult life, and evaluated the role of the NK1 receptor of adult animals that had been subjected to neonatal trauma. Rats were injected with 5 µl of SP-SAP (Cat. #IT-07, 30 ng/µl, 100 ng/µl, or 300 ng/µl) into the intrathecal space. Blank-SAP (Cat. #IT-21) was used as a control. The results indicate both that injury effects are isolated in the injured limb, and NK1 receptor-expressing cells are involved in processing this pain.
Siva AC, Wild MA, Kirkland RE, Nolan MJ, Lin B, Maruyama T, Yantiri-Wernimont F, Frederickson S, Bowdish KS, Xin H (2008) Targeting CUB domain-containing protein 1 with a monoclonal antibody inhibits metastasis in a prostate cancer model. Cancer Res 68:3759-3766. doi: 10.1158/0008-5472.CAN-07-1657
Summary: CUB domain-containing protein 1 (CDCP1) is an antigen expressed on several metastatic cancers, as well as on CD34+ and CD133+ myeloid leukemic blast cells. After demonstrating in vitro activity of the monoclonal antibody 25A11 with Mab-ZAP (Cat. #IT-04) and Hum-ZAP (Cat. #IT-22) the authors had a custom conjugation of 25A11 and saporin made for testing in mice. Goat-IgG-SAP (Cat. #IT-19) was used as a control for in vivo experiments, and saporin (Cat. #PR-01) was the control in vitro. The direct conjugate significantly inhibited tumor growth as well as metastasis in vivo.
Kline IV RH, Wiley RG (2008) Spinal mu-opioid receptor-expressing dorsal horn neurons: role in nociception and morphine antinociception. J Neurosci 28:904-913. doi: 10.1523/JNEUROSCI.4452-07.2008
Summary: The authors used Dermorphin-SAP (Cat. #IT-12) to investigate the function of spinal cord mu-opioid receptor (MOR)-expressing dorsal horn neurons in nociception and morphine analgesia. Rats were treated with 500 ng intrathecal injections of Dermorphin-SAP; 500 ng of Blank-SAP (Cat. #IT-21), and up to 1 µg of Saporin (Cat. #PR-01) were used as controls. The data indicate that MOR-expressing dorsal horn neurons are necessary for morphine action and play a role in nocifensive responses to persistent pain in the formalin test.
Berdiev RK, Chepurnov SA, Veening JG, Chepurnova NE, van Luijtelaar G (2007) The role of the nucleus basalis of Meynert and reticular thalamic nucleus in pathogenesis of genetically determined absence epilepsy in rats: A lesion study. Brain Res 1185:266-274. doi: 10.1016/j.brainres.2007.09.010
Summary: Absence seizures due to epilepsy usually occur during passive behavior. This work investigated the role of the cholinergic nucleus basalis of Meynert (NB) and the reticular thalamic nucleus (RT) in these seizures. Rats received either 75 ng of 192-IgG-SAP (Cat. #IT-01) or the control, mouse IgG-SAP (Cat. #IT-18), into the NB and the RT. Loss of cholinergic neurons in the NB resulted in an increased number of spike-and-wave discharges, a marker for absence seizures.
Shank EJ, Seitz PK, Bubar MJ, Stutz SJ, Cunningham KA (2007) Selective ablation of GABA neurons in the ventral tegmental area increases spontaneous locomotor activity. Behav Neurosci 121:1224-1233. doi: 10.1037/0735-7044.121.6.1224
Summary: To further examine the importance of the ventral tegmental area (VTA) g-aminobutyric acid (GABA) neurons in behavioral function, the authors lesioned the VTA of rats with dermorphin-saporin (Cat. #IT-12). Lesioned animals received 1 or 2 pmol/200 nl bilateral injections of conjugate; blank-SAP (Cat. #IT-21) was used as a control. Rats treated with dermorphin-SAP displayed significantly elevated motility as compared to control animals. Rats receiving 1 pmol of dermorphin-SAP returned to normal motility 14 days after treatment, but rats receiving 2 pmol maintained the increased motility through day 14.
Likhtik E, Popa D, Apergis-Schoute J, Fidacaro GA, Pare D (2007) Lesion of intercalated (ITC) amygdala neurons interfere with extinction of classically conditioned fear responses. Neuroscience 2007 Abstracts 426.6/HHH29. Society for Neuroscience, San Diego, CA.
Summary: The acquisition of conditioned fear responses (CRs) is thought to involve the potentiation of synapses conveying information about the conditioned stimulus (CS) to the basolateral (BLA) amygdala. Expression of CRs would depend on the transfer of potentiated CS inputs by the BLA to the central amygdala (CE). In contrast, the mechanisms of extinction remain controversial. It has been proposed that ITC neurons, which receive BLA inputs and generate feedforward inhibition in CE, are in a key position to mediate extinction. In this view, NMDA-dependent potentiation of BLA inputs to ITC cells during extinction training, would dampen the impact of CS-related BLA activity on CE neurons, inhibiting CRs. However, this idea is difficult to test because ITC cells occur in small, lateromedially dispersed clusters, making conventional lesioning methods inadequate. Here, we took advantage of the fact that, compared to the rest of the amygdala, ITC cells express a much higher concentration of mu opioid receptors (muORs). As a result, we could lesion them by performing local injections of a peptide-toxin conjugate (demorphin conjugated to saporin, D-Sap) that selectively targets cells expressing muORs. Control rats received injections of saporin conjugated to a blank peptide (B-Sap). On Day 1, intact rats were subjected to a standard cued fear conditioning protocol in context A. On Day 2, they received 20 CS alone presentations in a different context (B). On Day 3, rats then received either D-Sap or B-Sap injections in the ITC cell masses. One week later, extinction recall was tested in context B with 10 CS alone presentations. Compared to control (B-Sap) rats (n=10), ITC-lesioned rats (n=5) had an extinction deficit (ANOVA, F=11.687, p = 0.005). Post-hoc t-tests comparing % time freezing during the first five or last five CSs revealed that rats with ITC lesions had significantly higher freezing levels throughout the extinction recall test (p<0.002 for both tests). These differences were not attributable to a non-specific increase in freezing or anxiety levels as exploratory behaviors in a novel open field in control and ITC-lesioned rats were indistinguishable. Overall, these results indicate that ITC cells are involved in the expression of extinction.
Li A-J, Dinh TT, Ritter S (2007) Destruction of NPY receptor expressing neurons in the arcuate nucleus causes obesity and hyperphagia without increasing lateral hypothalamic orexigenic peptide gene expression. Neuroscience 2007 Abstracts 524.20/BBB20. Society for Neuroscience, San Diego, CA.
Summary: NPY-SAP, a conjugate of neuropeptide Y (NPY) and the ribosomal inactivating toxin, saporin (SAP), is a compound that selectively lesions NPY receptor-expressing neurons. Previously we showed that injection of NPY-SAP into the hypothalamic arcuate nucleus (ARC) induces hyperphagia and obesity in rats. To further investigate the mechanisms responsible for NPY-SAP-induced obesity, we injected NPY-SAP or blank-saporin (B-SAP) control into the ARC and subsequently examined the expression of two orexigenic neuropeptide genes in the lateral hypothalamic area (LHA), which is densely innervated by ARC neurons. Our hypothesis was that loss of leptin-sensitive neurons in the ARC in the NPY-SAP injected rats would lead to increased expression of orexigenic neurons elsewhere in the hypothalamic feeding circuitry. Body weight gain and food intake were dramatically increased in the NPY-SAP group. In addition, expression of NPY and cocaine- and amphetamine-regulated transcript (CART) mRNA was significantly reduced in the ARC of obese rats, indicating a loss of NPY receptor-expressing NPY and CART neurons in this region. In contrast, NPY and CART gene expression in the dorsomedial hypothalamic nucleus was unchanged in NPY-SAP rats, indicating that the NPY-SAP-induced lesion was limited to the ARC. However, contrary to our hypothesis, expression of the orexigenic neuronpeptides, melanin-concentrating hormone (MCH) or prepro-orexin mRNA in LHA was not enhanced, but was slightly reduced in the NPY-SAP rats. These results indicate that an enhancement of MCH or orexin expression in the LHA is not necessary for the hyperphagia and obesity observed after NPY-SAP lesions in the ARC. Supported by PHS grant #DK 40498.
Dias MB, Nucci TB, Margatho LO, Antunes-Rodrigues J, Gargaglioni LH, Branco LG (2007) Raphe Magnus Nucleus is involved in ventilatory but not hypothermic response to CO2. J Appl Physiol 103(5):1780-1788. doi: 10.1152/japplphysiol.00424.2007
Summary: In this work the authors investigated the role that serotonergic neurons in the Raphe Magnus Nucleus (RMg) play in ventilatory and thermal responses to hypercapnia. 0.1 µl of 1 µM anti-SERT-SAP (Cat. #IT-23) was injected into the RMg of rats. Mouse IgG-SAP (Cat. #IT-18) was used as a control. Lesioned animals had a decreased ventilatory response to CO2, but hypercapnia-induced hypothermia was not affected. The data indicate that RMg serotonergic neurons contribute to CO2 ventilatory response but not to maintenance of ventilation.
Schiltz JC, Sawchenko PE (2007) Specificity and generality of the involvement of catecholaminergic afferents in hypothalamic responses to immune insults. J Comp Neurol 502:455-467. doi: 10.1002/cne.21329
Summary: Interleukin-1 (IL-1) is one of the cytokines that mediates interactions between the immune system and the central nervous system. 380-ng injections of anti-DBH-SAP (Cat. #IT-03) were made into the paraventricular nucleus (PVH) of rats. Saporin (Cat. #PR-01) and mouse IgG-SAP (Cat. #IT-18) were used as controls. Lesioned animals demonstrated reduced responses to administration of IL-1, but restraint stress responses were left intact. The data suggest that ascending catecholaminergic projections mediate PVH response to IL-1.
Truitt WA, Sajdyk TJ, Dietrich AD, Oberlin B, McDougle CJ, Shekhar A (2007) From anxiety to autism: spectrum of abnormal social behaviors modeled by progressive disruption of inhibitory neuronal function in the basolateral amygdala in Wistar rats. Psychopharmacology (Berl) 191(1):107-118. doi: 10.1007/s00213-006-0674-y
Summary: The amygdala has been identified as being involved in social behaviors. Six 4 ng injections of SSP-SAP (Cat. #IT-11) were administered bilaterally into the basolateral nucleus (BLA) of the amygdala of rats. Blank-SAP (Cat. #IT-21) was used as a control. Results of a social interaction paradigm suggest that in normal animals social inhibition can be overcome by habituation. In lesioned animals, however, social inhibition is not reversed by habituation, indicating that NK-1 receptor-expressing GABAergic interneurons in the BLA are important in this system.
Furlong T, Carrive P (2007) Neurotoxic lesions centered on the perifornical hypothalamus abolish the cardiovascular and behavioral responses of conditioned fear to context but not of restraint. Brain Res 1128(1):107-119. doi: 10.1016/j.brainres.2006.10.058
Summary: This work examined the role of orexin-containing neurons in the perifornical hypothalamus (PeF) during stress response. Orexin-SAP (Cat. #IT-20) or the control conjugate blank-SAP (Cat. #IT-21) was injected into the PeF of pre-conditioned rats. Tests measuring restraint and conditioned fear to context were then performed on the lesioned animals. While the lesioning was not specific enough to connect results to orexin-containing neurons, the data indicate that the PeF is critical for some forms of stress, but not others.
Masawaki A, Sugiyo S, Shimoda T, Sakai Y, Watanabe M, Moritani M, Yoshida A, Niwa H, Takemura M (2006) Formalin-induced pain-related responses in rat lacking neurokinin-1 receptor neurons in the trigeminal nucleus caudalis. Neuroscience 2006 Abstracts 50.8. Society for Neuroscience, Atlanta, GA.
Summary: This study examines the effect of intra cisterna magna injection of substance P (SP) conjugated to saporin (SP-Sap; 5µM, 5µl) on formalin-induced pain-related behavior (PRB; face scrubbing behavior ) and c-Fos expression in the trigeminal nucleus caudalis (SpVc). In SP-Sap-treated rats, the numbers of NK-1-immunoreactive neurons in lamina I of the SpVc decreased compared with those in saline- or blank Sap-treated rats. The mean numbers ±SEM of PRB /5 min at the first phase (0-5 min after For injection) were 58.2±19.2 in the SP-Sap-treated rats, 115.6±14.0 in the saline treated rats and 86.9±45.7 in the blank-Sap-treated rats. The numbers at the quiescent period (5-10 min) were 45.2±26.3 in the SP-Sap- treated rats, 93.6±26.5 in the saline treated rats and 69.4±16.3 in the blank-Sap-treated rats. These at the former second phase (10-50 min) were 58.1±22.3 in the SP-Sap-treated rats, 133.6±26.1 in the saline treated rats and 95.8±29.6in the blank-Sap-treated rats. These at the latter second phase (55-90 min) were 7.0±5.6 in the SP-Sap-treated rats,13.7±12.4 in the saline treated rats and 10.4±22.5 in the blank-Sap-treated rats. These results indicate that formalin-induced nociceptive responses in the SP-Sap-treated rats are reduced.
Bugarith K, Dinh TT, Li AJ, Speth RC, Ritter S (2006) Featured Article: Basomedial hypothalamic injections of neuropeptide Y conjugated to saporin selectively disrupt hypothalamic controls of food intake. Targeting Trends 7(4)
Nattie E, Li A (2006) Neurokinin-1 receptor expressing neurons in the ventral medulla are essential for normal central and peripheral chemoreception in the conscious rat. J Appl Physiol 101(6):1596-1606. doi: 10.1152/japplphysiol.00347.2006
Summary: All known chemoreceptor sites in the mammalian brainstem are rich in the neurokinin-1 receptor (NK1r). The authors ask if these cells scattered throughout the ventral medulla are involved in central and peripheral chemoreception. Rats received 250-280 ng of SSP-SAP (Cat. #IT-11) into the cisterna magna, mouse IgG-SAP (Cat. #IT-18) was used as a control. The results indicate that NK1r neurons in the ventral medulla are involved in both central and peripheral chemoreception, during both waking and sleep states.
Parikh V, Sarter M (2006) Cortical choline transporter function measured in vivo using choline-sensitive microelectrodes: clearance of endogenous and exogenous choline and effects of removal of cholinergic terminals. J Neurochem 97(2):488-503. doi: 10.1111/j.1471-4159.2006.03766.x
Summary: A major projection of brain attention systems passes through the cholinergic portion of the cortical mantle. The authors investigated the role of high-affinity choline transporters (CHT) in the clearance of exogenous choline, as well as choline from newly released acetylcholine. 0.085 µg of 192-IgG-SAP (Cat. #IT-01) was injected into each hemisphere of the basal forebrain of rats (mouse IgG-SAP, Cat. #IT-18, was used as a control). The results demonstrate that no matter the source, increases in choline concentrations are cleared by CHT’s.
Foehr ED, Lorente G, Kuo J, Ram R, Nikolich K, Urfer R (2006) Targeting of the receptor protein tyrosine phosphatase beta with a monoclonal antibody delays tumor growth in a glioblastoma model. Cancer Res 66(4):2271-2278. doi: 10.1158/0008-5472.CAN-05-1221
Summary: The receptor protein tyrosine phosphatase ß (RPTPß) is overexpressed in astrocytomas, and is a potential target for tumor therapy. After testing antibodies against an extracellular domain of RPTPß in vitro with Mab-ZAP (Cat. #IT-04), two custom conjugates, 7E4B11-SAP and 7A9B5-SAP, were created by Advanced Targeting Systems. The authors tested the custom conjugates, using anti-DAT-SAP (Cat. #IT-25) as a positive control, and mouse IgG-SAP (Cat. #IT-18) as a negative control. The 7E4B11-SAP conjugate displayed significant antitumor activity in mice engrafted with U87 glioma cells.
Li A, Nattie E (2006) Catecholamine neurones in rats modulate sleep, breathing, central chemoreception and breathing variability. J Physiol 570(Pt 2):385-396. doi: 10.1113/jphysiol.2005.099325
Summary: Brainstem catecholamine (CA) neurons are thought to modulate the processing of sensory information and participate in the control of breathing. Using a 5 µg injection of anti-DBH-SAP (Cat. #IT-03), or a control injection of mouse-IgG-SAP (Cat. #IT-18) into the fourth ventricle, the authors investigated breathing frequency and wakefulness. The results suggest that CA neurons promote wakefulness, participate in central respiratory chemoreception, stimulate breathing frequency, and minimize breathing variability during REM sleep.
Bugarith K, Dinh TT, Li AJ, Speth RC, Ritter S (2005) Basomedial hypothalamic injections of neuropeptide Y conjugated to saporin selectively disrupt hypothalamic controls of food intake. Endocrinology 146(3):1179-1191. doi: 10.1210/en.2004-1166
Summary: The authors examined the effect of 48 ng injections of NPY-SAP (Cat. #IT-28) into the basomedial hypothalamus (BMH) on glucoprivic feeding in rats. While there was no evidence of retrograde transport, the lesions inhibited responses to intracerebroventricular leptin and ghrelin. Neither the feeding nor the hyperglycemic response to 2-deoxy-D-glucose was affected by the lesion, indicating that these hindbrain processes do not utilize neurons in the BMH. This work also describes dosing and injection parameter studies for the use of NPY-SAP.
Buffo A, Carulli D, Rossi F, Strata P (2003) Extrinsic regulation of injury/growth-related gene expression in the inferior olive of the adult rat. Eur J Neurosci 18(8):2146-2158. doi: 10.1046/j.1460-9568.2003.02940.x
Summary: Inferior olive (IO) cells of the CNS have the ability to regenerate axons after injury, even when the injury is close to the terminal field. After administration of 2.2 µg of 192-Saporin (Cat. #IT-01) and a control immunotoxin (mouse IgG-SAP, Cat #IT-18) to each ventricle in rats, two subsets of IO cells were discovered. Each subset responded differently to injury indicating that multiple mechanisms are responsible for their intrinsic regenerative potential.
Kanold PO, Kara P, Reid RC, Shatz CJ (2003) Role of subplate neurons in functional maturation of visual cortical columns. Science 301(5632):521-525. doi: 10.1126/science.1084152
Summary: Subplate neurons play a role in the development of connections between the thalamus and cerebral cortex. The authors used 0.5-µl injections of 0.25-1.0 mg/ml of ME20.4-SAP (Cat. #IT-15) to eliminate p75 receptor-positive neurons in the subplate of cats to investigate whether these neurons are involved in the organization and maturation of the visual cortex. This study also uses mouse IgG-saporin (Cat. #IT-18) as a control.
Lappi DA (2002) Featured Article: Control conjugates: The perfect companion for targeted toxins. Targeting Trends 3(1)
Tazzari PL, Bolognesi A, De Totero D, Falini B, Lemoli RM, Soria MR, Pileri S, Gobbi M, Stein H, Flenghi L, Martelli MF, Stirpe F (1992) Ber-H2 (anti-CD30)-saporin immunotoxin: a new tool for the treatment of Hodgkin's disease and CD30+ lymphoma: in vitro evaluation. Brit J Haemat 81:203-211. doi: 10.1111/j.1365-2141.1992.tb08208.x
Dinota A, Tazzari PL, Michieli M, Visani G, Gobbi M, Bontadini A, Tassi C, Fanin R, Damiani D, Grandi M, Pileri S, Bolognesi A, Stirpe F, Baccarani M, Tsuruo T, Tura S (1990) In vitro bone marrow purging of multidrug-resistant cells with a mouse monoclonal antibody directed against Mr 170,000 glycoprotein and a saporin-conjugated anti-mouse antibody. Cancer Res 50:4291-4294.