Internalization of (125I) alpha-bungarotoxin into rat suprachiasmatic nucleus neurons and dendrites

The distribution and rate of internalization of (125I) alpha-bungarotoxin (alpha-BuTX) was evaluated by quantitative electron microscopic autoradiography in rat suprachiasmatic nucleus. The ultrastructural distribution of silver grains was studied by both line source and 50% probability circle analyses. Line source analysis demonstrated that with increasing time following intracerebroventricular infusion, ligand is internalized from neuronal membranes. At all time points, real grain distribution was significantly different from randomly generated hypothetical grains. Quantification of silver grain localization by probability circle analysis indicated that membrane bound sources were mainly associated with axo-dendritic appositions, regardless of the length of time the tissue was exposed to radioligand. The compartment containing synaptic terminals was the most enriched when comparing real to hypothetical grains. By eight hours after intracerebroventricular infusion of specifically labelled alpha-BuTX binding sites were most likely to be within neurons and dendrites. These studies demonstrate that the majority of alpha-BuTX binding sites remain membrane bound with respect to time and may be associated with synaptic transmission; a significant proportion of silver grains are internalized within SCN neurons and dendrites.