Chapter 25 Immunolesion by 192IgG-saporin of rat basal forebrain cholinergic system: a useful tool to produce cortical cholinergic dysfunction

Abstract

Cholinergic lesion paradigms have been used to study the role of the cholinergic system in cortical arousal and cognitive function, and its implication in cognitive deficits that occur in Alzheimer's disease. In the last few years an increasing number of studies have applied neurotoxins including excitotoxins or cholinotoxins (e.g. AF64A) by stereotaxic injection into the Nbm to produce reductions in cortical cholinergic activity. One of the most serious limitations of these lesion paradigms is the fact that basal forebrain cholinergic neurons are always intermingled with populations of noncholinergic cells and that the cytotoxins used are far from being selective to cholinergic cells. Excitoxins when infused directly into the Nbm destroy non-specifically cell bodies but spare axons passing the injection site, whereas the specificity of AF64A to destroy cholinergic neurons depends on both the dosage applied and the site of injection. Recently, a monoclonal antibody to the low-affinity nerve growth factor (NGF) receptor, 192IgG, coupled to a cytotoxin, saporin, has been described as an efficient and selective immunotoxin for the NGF-receptor bearing cholinergic neurons in rat basal forebrain. Intraventricular administration of the 192IgG-saporin conjugate appears to induce a nearly complete and specific lesion of neocortical and hippocampal cholinergic afferents. Other neuronal systems in the basal forebrain are spared by the immunotoxin. Electrolytic, ibotenic acid, and cholinergic immunotoxic lesions of cholinergic basal forebrain nuclei resulted in slightly different effects on cortical cholinergic markers: Electrolytic lesion of the Nbm did not change M1-mAChR but resulted in reduced M2-mAChR in frontal and parietal cortices 1 week after lesion. Ibotenic acid lesion of the nucleus basalis did not alter M1-mAChR in any cortical region but led to enhanced M2-mAChR binding in the parietal cortex only. When applying the cholinergic immunotoxin 192IgG-saporin, both M1- and M2-mAChR binding sites were increased in a number of cortical areas 1 week after lesion. This comparison suggests that possibly the destruction of non-cholinergic basal forebrain cells by ibotenic acid and electrolytic lesion, might partly contribute to these different cortical effects. NMDA receptor binding was markedly reduced and AMPA, kainate, and GABAA receptor binding has been significantly increased in cortical regions displaying a reduced activity of AChE and decreased levels of high-affinity choline uptake sites due to immunolesion of the basal forebrain cholinergic system. Equivalent changes in cortical glutamate and GABA receptor subtype levels have been observed 7 days after electrolytic or ibotenic acid lesion of the Nbm. The data suggest that cholinergic immunolesion by 192IgG-saporin exhibits a valuable tool to produce specific cholinergic deficits in rats, which can be used as a model to study the effect of treatment with various drugs for compensating the impaired cortical cholinergic input.