Synaptic junctions isolated from cerebellum and forebrain: Comparisons of morphological and molecular properties

Abstract

Synaptic junction (SJ) fractions have been isolated from rat cerebellum which are similar to forebrain SJs on the basis of morphology and enrichment of synaptic structures. The polypeptide compositions of SJ fractions were analyzed by one- and two-dimensional SDS-gel electrophoresis and stained by the silver technique. Equivalent numbers of proteins that possess similar relative mobilities (Mr), isoelectric points and staining intensities were present in cerebellum and forebrain synaptic fractions. A few prominent differences were observed between cerebellum and forebrain synaptic fractions; cerebellum SJs contained a 240 K protein that was not detected in the forebrain and the 52,000 K, major PSDp protein was present in forebrain SJs in amounts that are approximately 5-fold greater than in cerebellum SJ fractions. The identity of the cerebellum mPSDp was verified by electrophoretic mobility, peptide fingerprinting and [125I]calmodulin binding. Differences between various synaptic fractions in mannose containing glycoproteins were examined by the binding of [125I]concanavalin A (Con A) to gels. On the basis of apparent molecular weights, the glycoproteins in forebrain and cerebellum SPMs were very similar. In contrast, however, the prominent glycoproteins that reside in the postsynaptic junctional membrane of forebrain SJs were undetectable in SJ fractions isolated from cerebellum. SJ fractions from cerebellum contained their own distinct group of Con A binding glycoproteins. SPM and SJ fractions from forebrain and cerebellum were examined for receptors for excitatory (aspartate, glutamate and kainic acid) and inhibitory (GABA) neurotransmitters and the benzodiazepine analog flunitrazepam. On the basis of relative receptor contents, SJ fractions isolated from either brain region were qualitatively similar and bound significant amounts of excitatory and inhibitory transmitters. These findings support the notion that SJs from cerebellum contain a distinct class of synaptic elements that are in large part derived from asymmetric, type I synapses.