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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