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Ultrastructural distribution of alpha-bungarotoxin binding sites in the suprachiasmatic nucleus of the rat hypothalamus
Authors:M. M. Miller  R. B. Billiar  A. Beaudet
Affiliation:(1) Departments of Obstetrics and Gynecology, Royal Victoria Hospital, Montreal, Canada;(2) Department of Experimental Medicine, McGill University Faculty of Medicine and Center for the Study of Reproduction, Montreal, Canada;(3) Department of Neurology and Neurosurgery, Montreal Neurological Institute, Montreal, Canada;(4) Department of Obstetrics and Gynecology, Royal Victoria Hospital, Women's Pavilion, 687 Pine Avenue West, H3A 1A1 Montreal, PQ, Canada
Abstract:Summary The distribution of (125I) alpha bungarotoxin (agr-BTX) binding sites in the suprachiasmatic nucleus (SCN) of the adult female rat was examined by electron-microscopic autoradiography. The ultrastructural distribution of silver grains was analysed by line source, direct point count, and 50% probability circle methods. Real grain distribution was significantly different from that of randomly generated hypothetical grains. Line source analysis demonstrated two populations of sources: one associated with membranes, and one inside neuronal structures. Probability circle analysis of shared grains indicated that membrane-bound-radioactive sources were mainly asssociated with axo-dendritic appositions. Only a small proportion of labeled neuronal interfaces exhibited synaptic differentiations in the plane of section. However, the compartment containing synaptic terminals was the most enriched when comparing real to hypothetical grains. Probability circle analysis of exclusive grains demonstrated that sources that were not associated with neuronal plasma membranes were likely to be within nerve cell bodies and dendrites. It is concluded that the majority of specifically labeled agr-BTX binding sites in the SCN is membrane bound, and may be associated with axodendritic synaptic transmission. The presence of a significant proportion of the label in the soma and dendrites of suprachiasmatic neurons 24 h after ventricular infusion suggests that some of the labeled binding sites (junctional or nonjunctional) may be internalized within these two compartments.
Keywords:Alpha-bungarotoxin  Autoradiography  Cholinergic receptors  Electron microscopy  Suprachiasmatic nucleus  Rat
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