References

Hess PR, Buntzman AS, Murray SL, Young EF, Frelinger JA (2009) Featured Article: Selective deletion of CD8+ T cells by saporin-coupled MHC class I tetramers. Targeting Trends 10(1)

Related Products: Streptavidin-ZAP (Cat. #IT-27), Saporin Goat Polyclonal, affinity-purified FITC-labeled (Cat. #AB-15APFL)

Read the featured article in Targeting Trends.

See Also:

• Hess PR et al. Selective deletion of antigen-specific CD8+ T cells by MHC class I tetramers coupled to the type I ribosome-inactivating protein saporin. Blood 109:3300-3307, 2007.

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.

Related Products: 192-IgG-SAP (Cat. #IT-01), Mouse IgG-SAP (Cat. #IT-18)

Anderberg C, Li H, Fredriksson L, Andrae J, Betsholtz C, Li X, Eriksson U, Pietras K (2009) Paracrine signaling by platelet-derived growth factor-CC promotes tumor growth by recruitment of cancer-associated fibroblasts. Cancer Res 69(1):369-378. doi: 10.1158/0008-5472.CAN-08-2724 PMID: 19118022

Usage: Receptor stimulation and immunoprecipitation.  B16/mock, B16/PDGF-C, and porcine aortic endothelial cells stably expressing PDGFR-a (a-PAE) were stimulated with 100 ng/mL core PDGF-CC for 1 h on ice. Immunoprecipitation was done with an antibody to fibroblast growth factor-2.  Western blot analysis was done with 1 ug/mL antibody.

Related Products: Fibroblast Growth Factor Rabbit Polyclonal, mammalian (Cat. #AB-07)

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.

Related Products: Dermorphin-SAP / MOR-SAP (Cat. #IT-12), Blank-SAP (Cat. #IT-21)

Burk JA, Lowder MW, Altemose KE (2008) Attentional demands for demonstrating deficits following intrabasalis infusions of 192 IgG-saporin. Behav Brain Res 195:231-238. doi: 10.1016/j.bbr.2008.09.006

Summary: Attentional processing has been shown to be dependent on basal forebrain cholinergic inputs to the cerebral cortex. In this work the authors wished to specify which components should be used to demonstrate deficits following the loss of these neurons. Rats received 200 ng intrabasalis infusions of 192-IgG-SAP (Cat. #IT-01). Testing of lesioned animals indicated that attentional deficits are due to increase of overall attentional task demands as opposed to any single task parameter.

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

Fitz NF, Gibbs RB, Johnson DA (2008) Selective lesion of septal cholinergic neurons in rats impairs acquisition of a delayed matching to position T-maze task by delaying the shift from a response to a place strategy. Brain Res Bull 77:356-360. doi: 10.1016/j.brainresbull.2008.08.016

Summary: It has been theorized that the effect of cholinergic lesions of the medial septum on learning depend on the stressful nature of the task being learned. The authors injected 0.2 µg of 192-IgG-SAP (Cat. #IT-01) into the medial septum of rats then examined the strategies used by these animals to learn a delayed matching to position T-maze task. Lesioned animals were less able to switch from one strategy to another, indicating that this mechanism is the primary one affected by septal cholinergic lesions.

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

Milstein JA, Dalley JW, Robbins TW (2010) Methylphenidate-induced impulsivity: pharmacological antagonism by beta-adrenoreceptor blockade. J Psychopharmacol 24:309-321. doi: 10.1177/0269881108098146

Summary: In this work bilateral 20 ng intracortical injections of anti-DBH-SAP (Cat. #IT-03) were used to examine the role of noradrenergic neurons in the control of psychostimulant-induced impulsivity. Although β-adrenoreceptor blockade abolished this impulsivity, lesioning noradrenergic neurons in the cortex had no effect. The data indicate that modulation of impulsive responding in this model is controlled by β-adrenoreceptors.

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

Zhang W, Gardell S, Zhang D, Xie JY, Agnes RS, Badghisi H, Hruby VJ, Rance N, Ossipov MH, Vanderah TW, Porreca F, Lai J (2009) Neuropathic pain is maintained by brainstem neurons co-expressing opioid and cholecystokinin receptors. Brain 132:778-787. doi: 10.1093/brain/awn330

Summary: It has been hypothesized that a subset of rostral ventromedial medulla (RVM) neurons co-expressing the cholecystokinin type 2 receptor and the mu-opioid receptor are responsible for the maintenance of neuropathic pain. Rats were treated with 50-ng bilateral RVM injections of Dermorphin-SAP (Cat. #IT-12), CCK-SAP (Cat. #IT-31), or saporin (Cat. #PR-01) as a control. Lesion of the RVM neurons prevented hyperalgesia in response to CCK treatment, and shortened abnormal pain states caused by sciatic nerve injury.

Related Products: Dermorphin-SAP / MOR-SAP (Cat. #IT-12), CCK-SAP (Cat. #IT-31), Saporin (Cat. #PR-01)

Laplante FP, Dufresne M, Lappi DA, Sullivan RM (2008) Depletion of cholinergic neurons in the nucleus accumbens and its possible involvement in schizophrenic symptomatology. Neuroscience 2008 Abstracts 761.18/FF34. Society for Neuroscience, Washington, DC.

Summary: Schizophrenia is a mental disorder characterized by dysfunctions in several neurotransmitter systems including the central cholinergic system. While alterations in cholinergic neurotransmission have been demonstrated in schizophrenic brains, their biological significance remains to be established. Post-mortem studies of schizophrenic patients have shown a reduction in the density of cholinergic interneurons in the striatum, most prominently in the ventral striatum or nucleus accumbens (N. Acc). Intra-accumbens acetylcholine interacts functionally with the mesolimbic dopaminergic system and is believed to dampen the effects of excessive dopamine (DA) release. Therefore, we hypothesize that a reduction in the density of cholinergic neurons in the N. Acc will be behaviorally relevant, if not causal, to the enhanced (ventral) striatal dopaminergic neurotransmission described in schizophrenia and may contribute substantially to the emergence of schizophrenic symptomatology. In this study we aimed to reproduce in rats a selective reduction in N.Acc. cholinergic cell density, and study the neurophysiological and behavioural consequences of these lesions, relevant to the neuropsychopathology of schizophrenia. A novel saporin immunotoxin coupled with an antibody targeting choline acetyltransferase (ChAT) has been developed. We microinjected this immunotoxin bilaterally (0.5 μg/μl; 0.5 μl) into the N. Acc (core and shell) of adult male Srpague-Dawley rats. Using immunohistochemistry to quantify ChAT staining, we have confirmed that this toxin caused a 40-50 % loss in the number of cholinergic neurons in this region within two weeks post-injection. Lesioned rats exhibited significantly higher spontaneous locomotor activity than control rats and were shown to be hypersensitive to the locomotor activating effects of amphetamine and quinpirole. Furthermore, in separate groups of animals, we have observed in lesioned rats, a reduction in the prepulse inhibition of the acoustic startle response. Taken together, it is proposed that reduction of cholinergic neurons in the N. Acc triggers an enhanced DA responsivity in the N.Acc which may prove highly effective in reproducing behavioral abnormalities analogous to those found in schizophrenia. The neurophysiological consequences of these lesions on DA neurotransmission will be further addressed by measuring both pre- and postsynaptic indices of DA function in this region.

Related Products: Anti-ChAT-SAP (Cat. #IT-42)

Datta S, Chatterjee K, Kline IV RH, Wiley RG (2008) Lumbar intrathecal CCK-saporin: anatomic and nociceptive effects. Neuroscience 2008 Abstracts 773.4/MM32. Society for Neuroscience, Washington, DC.

Summary: Lumbar intrathecal CCK (cholecystokinin) appears anti-opiate in nocifensive reflex testing and may be important in opiate-resistant neuropathic pain states suggesting a role for CCK receptor-expressing dorsal horn neurons in nociception. In the present study, we sought to determine if selective destruction of CCK receptor-expressing superficial dorsal horn neurons alters pain sensitivity or the analgesic potency of morphine using the targeted cytotoxic conjugate (CCK-sap) of CCK to saporin, a ribosome inactivating protein. 28 adult Sprague Dawley rats were injected via lumbar intrathecal catheter with CCK-sap in doses of 500 ng (n=2), 350 ng (n=3), 700 ng (n=3), 1000 ng (n=4), 1500 ng (n=4), or 3000 ng (n=4). Controls included PBS (n=4) or 1500 ng of plain, unconjugated saporin (n=4). 2 weeks later rats were sacrificed. Lumbar spinal cords were frozen sectioned at 40 µm. One-in-six series of transverse sections at L4-6 were immunostained for CCK. Two rats were injected with 1500 ng of CCK-sap followed by transcardiac aldehyde perfusion in 72 hours. L5 Dorsal root ganglia (DRG) sections were stained with cresyl violet and examined for signs of acute cytotoxicity (chromatolysis and karyohexis). 350 to 1500 ng of intrathecal CCK-sap were well tolerated with no obvious signs of any toxicity. 3000 ng of intrathecal saporin led to motor signs within 72 hours including increased muscle tone, leading to tonic hind limbs extension. Subsequently, twelve Long Evans female rats were tested before and after intrathecal injection of either PBS (n=8) or CCK-sap, 1500 ng (n=4) on: 1 – cold plate (15 °C); 2 – thermal preference shuttle box testing (15/45°C); 3 – hotplate at 44°C, 47°C and 52°C and 4 – thermal preference after morphine (0.5, 1 and 2.5 mg/kg s.c). Anatomical analysis revealed that 1500 ng of CCK-sap decreased CCK immunostaining in the L4-6 Dorsal horn. No acute cytotoxicity was seen in the DRG with1500 ng CCK-sap. Intrathecal CCK-sap was well tolerated at doses ≤1500 ng. CCK-sap produced increased hot side time and decreased crossovers in the thermal preference test. In contrast, CCK-sap decreased latency to first hindpaw lift and increased total responding on the 44 °C hotplate. CCK-sap rats also showed increased hot side time at 45° C at all morphine doses (0, 1 and 2.5 mg/kg s.c.) also with decreased crossovers. We interpret these observations to indicate that CCK-sap produced increased nocifensive reflex responding on the 44° C hotplate consistent with positive modulation of motor responsiveness, and CCK-sap reduced aversion to 45° C heat consistent with an analgesic effect that was additive with morphine.

Related Products: CCK-SAP (Cat. #IT-31)