Resolution of proteins from subfractions of nerve endings

W. J. McBride, James vanTassel

Research output: Contribution to journalArticle

13 Scopus citations

Abstract

An ultracentrifugation procedure is described for the isolation of ostomically sensitive nerve endings (synaptosomes) using an isotonic Ficoll-sucrose discontinuous gradient. These nerve endings wee lysed and their individual components isolated by isopycnic centrifugation in discontinuous sucrose gradients. With this procedures, highly purified fractions of nerve ending soluble material, synaptic vesicles, synaptic plasma membranes and nerve ending mitochondria were prepared. Polyacrylamide gel electrophoresis of the proteins from these subractions revealed that the soluble component had 19 detectable bands, the synaptic vesicles had 22 bands, the synaptic plasma membrane had 20 bands, and the mitochondria had 22 bands. The protein gel profile of the nerve ending mitochondria resembled the profile of the mitochondria obtained from the cell bodies of neurones and glia. Comparison of the profiles of the soluble proteins of the cell bodies (neuronal plus glial) and the nerve endings revealed one qualitative and 3 quantitative differences between the two. The synaptic vesicles had a complex gel pattern which did not resemble the pattern of any of the other components in the nerve endings. It was possible to obtain an insoluble fraction containing, in addition to large quantities of electron dense amorphous material, structures resembling synaptic complexes by treating the synaptic plasme membranes with Triton X-100. This insoluble fraction had a protein gel pattern consisting of 13 bands; 5 of these bands were not found in the protein profile of the soluble material.

Original languageEnglish (US)
Pages (from-to)177-187
Number of pages11
JournalBrain research
Volume44
Issue number1
DOIs
StatePublished - Sep 15 1972

ASJC Scopus subject areas

  • Neuroscience(all)
  • Molecular Biology
  • Clinical Neurology
  • Developmental Biology

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