ISOLATION AND PARTIAL CHARACTERIZATION OF RAT BRAIN SYNAPTIC PLASMA MEMBRANES

J. W. Gurd, L. R. Jones, H. R. Mahler, W. J. Moore

Research output: Contribution to journalArticle

285 Scopus citations

Abstract

Synaptic plasma membranes from the cortices of adult rat brain were isolated from synaptosomes prepared by flotation of a washed mitochondrial pellet (P2) in a discontinuous Ficoll sucrose gradient. Contamination of the synaptosome fraction by microsomes was estimated by enzymic and chemical analysis to be less than 15%. The purified synaptosome fraction was subjected to osmotic shock, subfractionated on a discontinuous sucrose gradient and the distribution of enzymic and chemical markers for synaptic plasma membranes, microsomal membranes and mitochondria was determined. Comparison of synaptosome subfractions prepared in the presence and absence of 1 mM NaH2 PO4/0.1 mM EDTA buffer pH 7.5, indicated that the ionic composition of the isolation medium markedly affected the distribution and enzymic composition of the subfractions. Synaptic plasma membranes prepared in the presence of PO4/EDTA exhibited a 10 fold enrichment in [Na+ + K+]ATPase and were characterized by less than 15 and 10% contamination by microsomes and mitochondria respectively. The polypeptide composition of the purified synaptic plasma membranes was compared with the microsomes and mitochondria by polyacrylamide gel electrophoresis in sodium dodecyl sulphate. No differences between the protein and glycoprotein composition of the synaptic plasma membranes and microsomes were detected. The mitochondria, in contrast, possessed a unique protein composition.

Original languageEnglish (US)
Pages (from-to)281-290
Number of pages10
JournalJournal of Neurochemistry
Volume22
Issue number2
DOIs
StatePublished - Feb 1974

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

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