Distribution and co-localization of choline acetyltransferase and p75 neurotrophin receptors in the sheep basal forebrain: implications for the use of a specific cholinergic immunotoxin

• Published in: Neuroscience 2001-01, Vol.104 (2), p.419-439
• Main Authors: Ferreira, G, Meurisse, M, Tillet, Y, Lévy, F
• Format: Article
• Language: English
• Subjects: acetylcholine; Animals; Antibody Specificity; Antineoplastic Agents, Phytogenic - toxicity; Basal Nucleus of Meynert - cytology; Basal Nucleus of Meynert - drug effects; Basal Nucleus of Meynert - metabolism; Biological and medical sciences; Cell Count; Central nervous system; Central neurotransmission. Neuromudulation. Pathways and receptors; Choline O-Acetyltransferase - drug effects; Choline O-Acetyltransferase - metabolism; Cholinergic Fibers - drug effects; Cholinergic Fibers - metabolism; Cholinergic Fibers - ultrastructure; Computer Science; Denervation - methods; Efferent Pathways - cytology; Efferent Pathways - drug effects; Efferent Pathways - metabolism; Female; Fundamental and applied biological sciences. Psychology; Immunoglobulin G - toxicity; Immunohistochemistry; immunolesion; Immunotoxins - toxicity; Life Sciences; Memory - drug effects; Memory - physiology; N-Glycosyl Hydrolases; Nerve Degeneration - chemically induced; Nerve Degeneration - metabolism; Nerve Degeneration - physiopathology; Plant Proteins - toxicity; Purkinje Cells - cytology; Purkinje Cells - drug effects; Purkinje Cells - metabolism; Receptor, Nerve Growth Factor - drug effects; Receptor, Nerve Growth Factor - metabolism; Ribosome Inactivating Proteins, Type 1; saporin; Saporins; Sheep; ungulates; Vertebrates: nervous system and sense organs
• ISSN: 0306-4522; 1873-7544
• DOI: 10.1016/S0306-4522(01)00075-6

The basal forebrain cholinergic system is involved in different forms of memory. To study its role in social memory in sheep, an immunotoxin, ME20.4 immunoglobulin G (IgG)–saporin, was developed that is specific to basal forebrain cholinergic neurons bearing the p75 neurotrophin receptor. The distribution of sheep cholinergic neurons was mapped with an antibody against choline acetyltransferase. To assess the localization of the p75 receptor on basal forebrain cholinergic neurons, the distribution of p75 receptor-immunoreactive neurons with ME20.4 IgG was examined, and a double-labeling study with antibodies against choline acetyltransferase and p75 receptor was undertaken. The loss of basal forebrain cholinergic neurons and acetylcholinesterase fibers in basal forebrain projection areas was assessed in ewes that had received intracerebroventricular injections of the immunotoxin (50, 100 or 150 μg) alone, as well as, in some of the ewes treated with the highest dose, with bilateral immunotoxin injections in the nucleus basalis (11 μg/side). Results indicated that choline acetyltransferase- and p75 receptor-immunoreactive cells had similar distributions in the medial septum, the vertical and horizontal limbs of the band of Broca, and the nucleus basalis. The double-labeling procedure revealed that 100% of the cholinergic neurons are also p75 receptor positive in the medial septum and in the vertical and horizontal limbs of the band of Broca, and 82% in the nucleus basalis. Moreover, 100% of the p75 receptor-immunoreactive cells of these four nuclei were cholinergic. Combined immunotoxin injections into ventricles and the nucleus basalis produced a near complete loss (80–95%) of basal forebrain cholinergic neurons and acetylcholinesterase-positive fibers in the hippocampus, olfactory bulb and entorhinal cortex. This study provides the first anatomical data concerning the basal forebrain cholinergic system in ungulates. The availability of a selective cholinergic immunotoxin effective in sheep provides a new tool to probe the involvement of basal forebrain cholinergic neurons in cognitive processes in this species.