Isolation and partial characterization of rat brain synaptic plasma membranes

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

Research output: Contribution to journalArticle

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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
Pages (from-to)281-290
Number of pages10
JournalJournal of Neurochemistry
Volume22
Issue number2
StatePublished - 1974

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Synaptic Membranes
Cell membranes
Synaptosomes
Rats
Brain
Microsomes
Mitochondria
Cell Membrane
Chemical analysis
Edetic Acid
Sucrose
Contamination
Ficoll
Osmotic Pressure
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
Flotation
Electrophoresis
Glycoproteins
Buffers

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Isolation and partial characterization of rat brain synaptic plasma membranes. / Gurd, J. W.; Jones, Larry; Mahler, H. R.; Moore, W. J.

In: Journal of Neurochemistry, Vol. 22, No. 2, 1974, p. 281-290.

Research output: Contribution to journalArticle

Gurd, J. W. ; Jones, Larry ; Mahler, H. R. ; Moore, W. J. / Isolation and partial characterization of rat brain synaptic plasma membranes. In: Journal of Neurochemistry. 1974 ; Vol. 22, No. 2. pp. 281-290.
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