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Synaptosomal and vesicular accumulation of L-glutamate, L-aspartate and D-aspartate.

Abstract：

:We examined the vesicular accumulation of the excitatory amino-acid (EAA) neurotransmitters, L-glutamate and L-aspartate, together with the non-metabolisable EAA analogue D-aspartate. Synaptosomes derived from whole brain were incubated in various concentrations of [3H]-amino acids under conditions to facilitate vesicular turnover. Synaptosomes were then lysed in hypotonic medium and vesicles immunoprecipitated with monoclonal anti-synaptophysin antibodies coupled to sepharose beads. Using this method, saturable vesicular accumulation was observed for [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate but not for the excitatory amino acid receptor ligands [3H]-AMPA or [3H]-kainate. Vesicular accumulation (t(1/2)=7.45 min) was markedly slower than synaptosomal accumulation (t(1/2)=1.03 min) and was substantially reduced at 4 degrees C. Maximal accumulation of [3H]-L-glutamate, [3H]-L-aspartate, and [3H]-D-aspartate was estimated to be 98, 68, and 112 pmol/mg of synaptosomal protein, respectively, and uptake affinities 1.6, 3.4, and 2.1 mM, respectively. Maximal accumulation of [3H]-L-glutamate was non-competitively inhibited by both 100 microM unlabeled L-aspartate and 100 microM D-aspartate, suggesting that all are accumulated into a common vesicular pool by different transporters.

journal_name

Neurochem Int

journal_title

Neurochemistry international

authors

Fleck MW,Barrionuevo G,Palmer AM

doi

10.1016/s0197-0186(01)00018-3

subject

Has Abstract

pub_date

2001-09-01 00:00:00

pages

217-25

issue

3

eissn

0197-0186

issn

1872-9754

pii

S0197-0186(01)00018-3

journal_volume

39

pub_type

杂志文章

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