References

Roberts RC, Harrison MB, Francis SMN, Wiley RG (1993) Differential effects of suicide transport lesions of the striatonigral or striatopallidal pathways on subsets of striatal neurons. Exp Neurol 124:242-252. doi: 10.1006/exnr.1993.1194

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Picklo MJ, Wiley RG, Lappi DA, Robertson D (1993) Noradrenergic lesioning using anti-DBH immunotoxin. Neuroscience 1993 Abstracts 771.10. Society for Neuroscience, Washington, DC.

Summary: Current methods of lesioning noradrenergic neurons have significant drawbacks due either to incompleteness, partial specificity, or reversibility. We sought to determine if an immunotoxin to dopamine ß-hydroxylase (DBH) would efficiently destroy noradrenergic neurons in vivo. The MAB3O8 monoclonal antibody to bovine DBH was obtained from Chemicon International (Temecula, CA). Antibody was disulfide coupled to the ribosome inactivating protein, saporin, using SPDP. The resulting immunotoxin was injected into anesthetized adult, male Sprague-Dawley rats. Injection sites in individual animals were the submandibular gland (3.1-12.5 µg), intravenous (6-31 µg), and intraventricular (1.8-4.8 µg). Three days after systemic injections, rats were reanesthetized and transcardially perfused with aldehyde fixative. Frozen sections of peripheral ganglia were processed for Nissi staining. In sections from sympathetic ganglia, most neurons showed severe chromatolysis characteristic of the cytotoxic effect of immunotoxins containing saporin. Sensory ganglia showed small numbers of similarly poisoned neurons. Eleven days after intraventricular injections, rats were sacrificed and brain sections stained either with cresyl violet (N¡ssl) or for tyrosine hydroxylase (TH) using indirect immunoperoxidase technique. Nissi staining of the locus coeruleus showed a decrease in the numbers of neurons as was confirmed by staining for TH. Dopaminergic neurons in the midbrain appeared unaffected as did catecholaminergic neurons in the caudal brainstem. We conclude that anti-DBH-saporin efficiently destroys noradrenergic neurons in the CNS and PNS. This immunotoxin may be a valuable lesioning tool with greater selectivity than has been previously available.

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Leanza G, Nilsson O, Wiley RG, Bjorklund A (1993) Dose-related effects of 192 IgG-saporin on basal forebrain cholinergic neurons: Biochemical, immunocytochemical and behavioral studies on neonatal and adult rats. Neuroscience 1993 Abstracts 771.12.

Summary: Highly selective and efficient depletions of the NGF receptor-positive cholinergic neurons in the basal forebrain, associated with profound behavioral changes, have been reported after treatment with 192 IgG-saporin, a recently introduced immunotoxin. In the present study, the effects of different doses of the toxin conjugate were compared following bilateral injections in the lateral ventricles of infant (2-3 days post-natal, 0.2 to 0.8 µg dose-range) and adult rats (1.25 to 10 µg dose-range). Five to eight weeks post-injection, dose-related behavioral effects of the lesion were evaluated in water maze, passive avoidance and locomotor activity tests, followed by biochemical and histologic analyses performed on tissue specimens. In the adult rats, the immunotoxin produced severe dose-dependent cognitive deficits in water maze task performance and passive avoidance retention that correlated with decline in choline acetyltransferase (ChAT) activity in neocortical and hippocampal areas (up to 95% reduction) and depletion of NGF receptor-positive neurons in the septal-diagonal band area and nucleus basalis (>95%). Similar dose-related patterns of ChAT activity reduction and neuronal loss throughout the basal forebrain were seen in rats neonatałly injected with the immunotoxin. ChAT levels in other regions such as brainstem, cerebellum or the spinal cord were found largely unaffected in both adult and neonatałly lesioned rats. The present results confirm 192 IgG-saporin as a powerful tool to induce specific and robust cholinergic lesions either during development or adulthood and emphasize the crucial role of the forebrain cholinergic system in cognitive functions.

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Contestabile A, Stirpe F (1993) Ribosome-inactivating proteins from plants as agents for suicide transport and immunolesioning in the nervous system. Eur J Neurosci 5(10):1292-1301. doi: 10.1111/j.1460-9568.1993.tb00915.x

Benecke S, Ostermann-Latif C, Mader M, Schmidt B, Unger JW, Westarp ME, Felgenhauer K (1993) Antibodies raised against synthetic peptides react with choline acetyltransferase in various immunoassays and in immunohistochemistry. J Neurochem 61(3):804-811. doi: 10.1111/j.1471-4159.1993.tb03590.x PMID: 8360685

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Attila M, Salvadori S, Balboni G, Bryant SD, Lazarus LH (1993) Synthesis and receptor binding analysis of dermorphin hepta-, hexa- and pentapeptides. Int J Pept Prot Res 42:550-559. doi: 10.1111/j.1399-3011.1993.tb00363.x

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Mesner LD, Truman PA, Hockensmith JW (1993) DNA-dependent adenosinetriphosphatase A: immunoaffinity purification and characterization of immunological reagents. Biochemistry 32(30):7772-7778. doi: 10.1021/bi00081a024 PMID: 8394114

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Dabadie H, Geffard M (1993) Identification of tryptamine and tryptamine-serotonin neurons in the rat dorsal raphe nuclei using specific antibodies. Synapse 14(2):178-183. doi: 10.1002/syn.890140211 PMID: 7687387

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Kitchener PD, Wilson P, Snow PJ (1993) Selective labelling of primary sensory afferent terminals in lamina II of the dorsal horn by injection of Bandeiraea simplicifolia isolectin B4 into peripheral nerves. Neuroscience 54(2):545-551. doi: 10.1016/0306-4522(93)90274-j

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Shigemoto R, Nakaya Y, Nomura S, Ogawa-Meguro R, Ohishi H, Kaneko T, Nakanishi S, Mizuno N (1993) Immunocytochemical localization of rat substance P receptor in the striatum. Neurosci Lett 153(2):157-160. doi: 10.1016/0304-3940(93)90311-8 PMID: 8392153

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