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Yaksh T, Fisher C, Hockman T, Wiese A (2017) Current and future issues in the development of spinal agents for the management of pain. Curr Neuropharmacol 15:232-259.. doi: 10.2174/1570159×14666160307145542

Summary: Although conscious pain experience is driven by signals mediated supraspinally, the more high intensity pain generated by strong stimuli, tissue injury, and nerve injury is encoded at the spinal dorsal horn level. The control of pain signals at the spinal dorsal horn level is a tempting target for targeted pain therapy. This review discusses the potential targets for pain therapeutics in the spinal dorsal horn, and some of the spinal agents used to modulate pain transmission through that location. The use of SSP-SAP (Cat. #IT-11) is mentioned as a neurokinin-1 targeted molecule that can block some pain transmission.

Related Products: SSP-SAP (Cat. #IT-11)

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.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Mouse IgG-SAP (Cat. #IT-18)

Nam H, Kerman I (2016) A2 noradrenergic neurons regulate forced swim test immobility. Physiol Behav 165:339-349. doi: 10.1016/j.physbeh.2016.08.020

Summary: Wistar-Kyoto rats are often used as a model of depression, and exhibit high levels of immobility when subjected to a forced swim test (FST). Researchers discovered relative hyperactivation in the locus coeruleus of WKY rats compared to the genetically related Wistar rats when exposed to one- and two-day FSTs. Lesser activation of A2 noradrenergic cell group was seen by diminished levels of FOS after both days of the FST. A2 noradrenergic neurons of Winstar rats were lesioned by injecting 2.2 ug of Anti-DBH-SAP (Cat. #IT-03) into the nucleus tractus solitaris (NTS). Lesioned rats exhibited increased FST immobility on both days of the test, similar to natural WKY behavior in the same test. These data indicate that the A2 noradrenergic cell group regulates FST behavior and that its hypoactivation may contribute to the unique behavioral phenotype of WKY rats.

Related Products: Anti-DBH-SAP (Cat. #IT-03)

Nohara S, Kato K, Fujiwara D, Sakuragi N, Yanagihara K, Iwanuma Y, Kajiyama Y (2016) Aminopeptidase N (APN/CD13) as a target molecule for scirrhous gastric cancer. Clin Res Hepatol Gastroenterol 40:494-503. doi: 10.1016/j.clinre.2015.11.003

Summary: Scirrhous gastric cancer has the worst prognosis of gastric carcinoma, and treatment with standard cancer therapies has had minimal success. In this work the authors target CD13 as a marker for scirrhous gastric cancer. A gastric cancer cell line was challenged with a CD13 antibody coupled to Mab-ZAP (Cat. #IT-04) in an in vitro cytotoxicity assay. The anti-CD13 complex was more cytotoxic than an anti-EpCAM-immmunotoxin. These data, combined with flow cytometry analysis and enzyme activity assays, demonstrate the expression of CD13 as a marker for scirrhous gastric cancer.

Related Products: Mab-ZAP (Cat. #IT-04)

Coradazzi M, Gulino R, Fieramosca F, Falzacappa L, Riggi M, Leanza G (2016) Selective noradrenaline depletion impairs working memory and hippocampal neurogenesis. Neurobiol Aging 48:93-102. doi: 10.1016/j.neurobiolaging.2016.08.012

Summary: Neuronal loss in the locus coeruleus (LC) of Alzheimer’s patients is well known, but the contribution of LC-derived noradrenergic afferents to learning and memory function is unknown. To model noradrenergic neuron degeneration in the LC, rats were bilaterally injected directly into the LC with 0.2 ug of Anti-DBH-SAP (Cat. #IT-03). Lesioned and sham-lesioned animals were tested behaviorally and exhibited robust working memory deficits but lesioning did not affect reference memory. They concluded that ascending noradrenergic afferents might be involved in more complex aspects of working memory, possibly via newly generated progenitors in the hippocampus.

Related Products: Anti-DBH-SAP (Cat. #IT-03)

Araldi D, Ferrari L, Levine J (2016) Gi-protein-coupled 5-HT1B/D receptor agonist sumatriptan induces type I hyperalgesic priming. Pain 157:1773-1782. doi: 10.1097/j.pain.0000000000000581

Summary: The present study explored the possibility that, like MOR and A1-adenosine receptor agonists, triptans would also induce type II hyperalgesic priming. In addition, they explored the 5-HT receptor subtypes at which triptans act (5-HT1B, 5-HT1D and 5-HT7) to induce priming. They report that while sumatriptan, a prototypical 5-HT1B/D receptor agonist induces hyperalgesic priming, this priming meets the criteria for type I rather than type II priming. Isolectin B4 (IB4)-saporin (Cat. #IT-10), was diluted in saline, and a dose of 3.2 μg, in a volume of 20 μL was administered intrathecally to rats. The neurotoxin [Sar9,Met(O2) 11]-substance P-saporin (SSP-Saporin, Cat. #IT-11) was diluted in saline, and a dose of 100 ng, in a volume of 20 μL was administered intrathecally. In a model of pain chronification, sumatriptan induces both mechanical hyperalgesia at the site of injection and type I hyperalgesic priming, in nociceptors innervating the cutaneous injection site.

Related Products: IB4-SAP (Cat. #IT-10), SSP-SAP (Cat. #IT-11)

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.

Related Products: CRF-SAP (Cat. #IT-13), Blank-SAP (Cat. #IT-21)

Read the featured article in Targeting Trends.

Lim J, Stafford B, Nguyen P, Lien B, Wang C, Zukor K, He Z, Huberman A (2016) Neural activity promotes long-distance, target-specific regeneration of adult retinal axons. Nat Neurosci 19:1073-1084. doi: 10.1038/nn.4340 PMID: 27399843

Summary: Axons in the CNS fail to regenerate after injury. Scientists sought to identify strategies that would allow retinal ganglion cell (RGC) axons to regenerate in the eye-to-brain pathway, and if that was possible, whether the axons could reconnect with their correct targets and restore visual function. It was previously shown that increasing mTOR signaling could trigger RGC axon regeneration. Several conditions were tested, but combining increased mTOR signaling and then exposing mice to high-contrast visual stimulation daily for 3 weeks scientists after optic nerve crush resulted in long distance RGC axon regeneration, re-innervation of the brain and partial recovery of a subset of visual behaviors. A 1:1000 dilution of Anti-Melanopsin (Cat. #AB-N38) was used for the immunohistochemical analysis of retinas, optic nerves and brain tissue.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

Gilabert-Oriol R, Thakur M, Haussmann K, Niesler N, Bhargava C, Görick C, Fuchs H, Weng A (2016) Saponins from Saponaria officinalis L. augment the efficacy of a Rituximab-immunotoxin. Planta Med 82:1525-1531. doi: 10.1055/s-0042-110495

Summary: It is known that triterpenoidal saponins that come from Saponaria officinalis, the plant that saporin comes from, increases the cytotoxicity of saporin by modulating its intracellular trafficking. Investigators wanted to know if this could increase the therapeutic affect of Rituximab-Saporin. In the presence of saponins, Rituximab-Saporin had a 700-fold increase in efficacy. Concentrations of 0.0001-1nM of Anti-CD22-SAP (Cat. #IT-37) and 0.001-10nM of Anti-CD25-SAP (Cat. #IT-24) were also tested in vitro with saponins for comparison. They saw a 170-fold and 25-fold increase in cytotoxicity, respectively. All conjugates were tested on Ramos cells, and differing levels of receptor expression could explain the drastic differences in cytotoxicity enhancement.

Related Products: Anti-CD22-SAP human (Cat. #IT-37), Anti-CD25-SAP human (Cat. #IT-24)

Palchaudhuri R, Saez B, Hoggatt J, Schajnovitz A, Sykes D, Tate T, Czechowicz A, Kfoury Y, Ruchika F, Rossi D, Verdine G, Mansour M, Scadden D (2016) Non-genotoxic conditioning for hematopoietic stem cell transplantation using a hematopoietic-cell-specific internalizing immunotoxin. Nat Biotechnol 34:738-745. doi: 10.1038/nbt.3584

Summary: To demonstrate correction of a clinically relevant disease, we employed CD45-SAP in a mouse model of sickle cell anemia and demonstrated our method achieved >90% donor cell chimerism, all mice in three groups (18/18), resulting in complete disease correction (red blood cell counts, hemoglobin levels, hematocrit levels and reticulocyte frequencies were returned to normal). If these pre-clinical results can be successfully translated to the clinic, it would greatly reduce conditioning-related toxicities and expand the use of hematopoietic stem cell transplantation.

Related Products: Streptavidin-ZAP (Cat. #IT-27)

Read the featured article in Targeting Trends.

Arroyo D, Kirkby L, Feller M (2016) Retinal waves modulate an intraretinal circuit of intrinsically photosensitive retinal ganglion cells. J Neurosci 36:6892-6905. doi: 10.1523/JNEUROSCI.0572-16.2016 PMID: 27358448

Summary: The researchers explore the neural circuits underlying the ipRGC driven light responses of the developing retina and the mechanisms by which retinal waves regulate these circuits. They demonstrate that, even in the presence of cholinergic waves, ipRGC gap junction microcircuits propagate light-driven signals, thus strongly contributing to the overall light response of the developing retina. Following fixation, retinas were washed in PBS and remounted onto a new piece of filter paper. They were incubated in blocking buffer and then in primary immunoreaction solution, 1:2500 rabbit anti-melanopsin (Cat. #AB-N38). Results show that, during development, ipRGCs form extensive gap junction microcircuits that shape the early retinal light response. Retinal waves exert a far-reaching, neuromodulatory influence on these circuits via dopaminergic modulation of gap junctions, thus potentially impacting the processing of early visual input.

Related Products: Melanopsin Rabbit Polyclonal (Cat. #AB-N38)

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.

Related Products: Oxytocin-SAP (Cat. #IT-46), Blank-SAP (Cat. #IT-21)

Mohammed M, Kulasekara K, Ootsuka Y, Blessing W (2016) Locus coeruleus noradrenergic innervation of the amygdala facilitates alerting-induced constriction of the rat tail artery. Am J Physiol Regul Integr Comp Physiol 310:R1109-1119. doi: 10.1152/ajpregu.00058.2016

Summary: The researchers tested the hypothesis that release of noradrenaline within the amygdala is important for the occurrence of SCVARS (sympathetic cutaneous vasoconstrictor alerting responses). A long-shanked 5-μl glass micropipette calibrated in 100-nl steps, was filled with vehicle or Anti-DBH-SAP (Cat. #IT-03). Anti-DBH-SAP (5 μg in 250 nl) or vehicle was injected into the amygdala during ∼1 min, and the pipette was left in place for an additional The locus coeruleus has been implicated in many aspects of emotional arousal, so that functional inhibition of the extensive locus coeruleus-derived noradrenergic innervation of centers known to be important in emotional arousal, including the amygdala, is likely to contribute to the therapeutic actions of clonidine-like agents. The locus coeruleus also has major reciprocal connections with the orexin-synthesizing neurons in the hypothalamus, and rats with genetically lesioned orexin receptor neurons (alternatively, oen could lesion with Orexin-SAP, Cat. #IT-20) have reduced emotional arousal as reflected in reduced SCVAR responses to alerting stimuli.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Orexin-B-SAP (Cat. #IT-20)

Dickey D, Thomas G, Dassie J, Giangrande P (2016) Method for confirming cytoplaintratumoral anti-HuD immunotoxinsmic delivery of RNA aptamers. (eds. Shum K, Rossi J). In: SiRNA Delivery Methods. Methods in Molecular Biology. 1364:209-217. Humana Press, New York, NY. doi: 10.1007/978-1-4939-3112-5_17

Objective: To describe a functional assay (RIP assay) to confirm cellular uptake and subsequent cytoplasmic release of an RNA aptamer which binds to a cell surface receptor expressed on prostate cancer cells (PSMA).

Summary: This publication details an in vitro functional assay to confirm that the aptamer retains function following conjugation to saporin and describe a cellular assay to measure aptamer-mediated saporin-induced cytotoxicity.

Usage: The folded biotinylated aptamer was mixed at a 1:4 molar ratio of Streptavidin-ZAP, confirmed by agarose gel, a PSMA enzymatic activity (NAALADase) assay performed. FGF-SAP was used as a control.

Related Products: Streptavidin-ZAP (Cat. #IT-27), FGF-SAP (Cat. #IT-38)

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.

Related Products: Oxytocin-SAP (Cat. #IT-46), Blank-SAP (Cat. #IT-21)

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

Related Products: Oxytocin-SAP (Cat. #IT-46), Blank-SAP (Cat. #IT-21)

Lee S, Diener K, Kaufman S, Krieger J, Pettersen K, Jejelava N, Arnold M, Watts A, Langhans W (2016) Limiting glucocorticoid secretion increases the anorexigenic property of Exendin-4. Mol Metab 5:552-565. doi: 10.1016/j.molmet.2016.04.008

Summary: Glucagon-like peptide-1 (GLP-1) analogs lower blood surgar levels and cause a loss of appetite. Exendin-4 (Ex-4) is a GLP-1 receptor agonist, and also increases glucocorticoid secretion. Several tests were conducted to determine if the released glucocorticoids interact with Ex-4’s anorexigneic effect. One method involved ablating hindbrain catecholaminergic neurons by stereotaxically injecting 42 ng of Anti-DBH-SAP (Cat. #IT-03) bilaterally into the paraventricular nucleus of the hypothalamus in rats. Animals were injected with equimolar concentrations of unconjugated Saporin (Cat. #PR-01) as a control. Anti-DBH-SAP lesions reduced the efficacy of Ex-4 to increase corticosterone secretion but increased the anorexigenic effect, indicating that Ex-4-dependent corticosterone secretion opposes Ex-4’s actions. Anti-DBH-SAP lesions increased Ex-4’s ability to reduce food intake and body weight.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Saporin (Cat. #PR-01)

Sedlik C, Heitzmann A, Viel S, Ait Sarkouh R, Batisse C, Schmidt F, De La Rochere P, Amzallag N, Osinaga E, Oppezzo P, Pritsch O, Sastre-Garau X, Hubert P, Amigorena S, Piaggio E (2016) Effective antitumor therapy based on a novel antibody-drug conjugate targeting the Tn carbohydrate antigen. Oncoimmunology 5:e1171434. doi: 10.1080/2162402X.2016.1171434

Summary: Scientists wanted to study the potential of Chi-Tn, a monoclonal antibody against a glycol-peptidic tumor-associated antigen, as an anticancer antibody-drug conjugate. They demonstrated that Chi-Tn specifically targeted tumor cells in vivo, using flow cytometry and deconvolution microscopy to show that Chi-Tn is rapidly internalized. Chi-Tn-SAP (ATS Custom Services) effectively killed Tn-positive cells, but had no effect on Tn-negative cells. Saporin (Cat. #PR-01) was used as control. The cytotoxicity of the Chi-Tn-SAP correlated with the level of tumoral Tn expression.

Related Products: Saporin (Cat. #PR-01), Custom Conjugates

Vierck C, Yezierski R, Wiley R (2016) Pain sensitivity following loss of cholinergic basal forebrain (CBF) neurons in the rat. Neuroscience 319:23-34. doi: 10.1016/j.neuroscience.2016.01.038

Objective: There is a large amount of research on the involvement of cholinergic mechanisms on spinal transmission of pain signals, indicating that cholinergic agonists can attenuate this kind of pain. In contrast, some studies have shown affective reactions to pain are suppressed by cholinergic antagonists. The authors investigated the disagreement between reflexive and affective reactions.

Summary: Lesioned rats displayed decreased escape from thermal stimulation, as well as loss of the normal hyperalgesic effect of sound stress. Results indicate that the basal forebrain cholinergic system plays a role in central processing of pain.

Usage: Administration of 192-IgG-SAP with a 4-μg injection into the left lateral ventricle of rats. Animals were tested in temperature escape and sound stress models.

Related Products: 192-IgG-SAP (Cat. #IT-01)

Schwartz M, Nguyen A, Warrier D, Palmerston J, Thomas A, Morairty S, Neylan T, Kilduff T (2016) Locus coeruleus and tuberomammillary nuclei ablations attenuate hypocretin/orexin antagonist-mediated rem sleep. eNeuro 3:ENEURO.0018-0016.2016. doi: 10.1523/ENEURO.0018-16.2016

Summary: To examine the mechanism by which the Orexin 1r/Orexin 2r antagonist almorexant decreases wakefulness and increases NREM and REM sleep the authors utilized Anti-DBH-SAP (Cat. #IT-03) and Orexin-B-SAP (Cat. #IT-20). Rats received 3-μg injections of Anti-DBH-SAP into the LC, or bilateral 57-80 ng injections of Orexin-SAP into the TMN. Both conjugates attenuated the increased REM sleep seen upon administration of almorexant without altering almorexant-induced changes in NREM sleep.

Related Products: Anti-DBH-SAP (Cat. #IT-03), Orexin-B-SAP (Cat. #IT-20)