Role of astrocytes in depolarization-coupled release of glutamate in cerebellar cultures.
Release of preloaded D-[3H]aspartate in response to depolarization induced by high potassium, N-methyl-D-aspartate (NMDA), alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) or the endogenous agonist glutamate was studied using cultured glutamatergic cerebellar granule neurons, cerebellar astrocytes, and corresponding cocultures. Release from the vesicular and the cytoplasmic glutamate pools, respectively, was distinguished employing the competitive, non-transportable glutamate transport inhibitor DL-threo-beta-benzyloxyaspartate (DL-TBOA). The results indicate that the release in response to AMPA (30 microM) in the presence of cyclothiazide (50 microM) to block desensitization, was of a vesicular origin. Pulses of 55 mM K+ caused a DL-TBOA resistant efflux of preloaded D-[3H]aspartate from astrocytes, indicating that this release was not mediated by glutamate transporters. The results furthermore support the notion of an important function of the astrocytes in the uptake of released glutamate, because DL-TBOA caused a large, apparent increase in the depolarization-coupled release of preloaded D-[3H]aspartate in the cocultures, compared to neuronal monocultures.
Department of Pharmacology, The Danish University of Pharmaceutical Sciences, DK-2100 Copenhagen, Denmark.
Lasse K
LK
Bak
Helle S
HS
Waagepetersen
Arne
A
Schousboe
D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism / Q000494:pharmacology; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects / Q000502:physiology*; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism*; D051379:Mice; D015739:Nocodazole / Q000494:pharmacology; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid / Q000494:pharmacology
C120673:benzyloxyaspartate; D001224:Aspartic Acid; D018698:Glutamic Acid; D018350:alpha-Amino-3-hydroxy-5-methyl-4-isoxazolepropionic Acid; D015739:Nocodazole