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Excitatory amino acid-stimulated uptake of 22Na+ in primary astrocyte cultures

HK Kimelberg, S Pang and DH Treble
Journal of Neuroscience 1 April 1989, 9 (4) 1141-1149; https://doi.org/10.1523/JNEUROSCI.09-04-01141.1989
HK Kimelberg
Department of Biochemistry, Albany Medical College, Albany, New York 12208.
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S Pang
Department of Biochemistry, Albany Medical College, Albany, New York 12208.
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DH Treble
Department of Biochemistry, Albany Medical College, Albany, New York 12208.
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Abstract

In this study we have found that L-glutamic acid, as well as being taken up by a Na+-dependent mechanism, will stimulate the uptake of 22Na+ by primary astrocyte cultures from rat brain in the presence of ouabain. By simultaneously measuring the uptake of 22Na+ and L-3H- glutamate a stoichiometry of 2–3 Na+ per glutamate was measured, implying electrogenic uptake. Increasing the medium K+ concentration to depolarize the cells inhibited L-3H-glutamate uptake, while calculations of the energetics of the observed L-3H-glutamate accumulation also supported an electrogenic mechanism of at least 2 Na+:1 glutamate. In contrast, kinetic analysis of the Na+ dependence of L-3H-glutamate uptake indicated a stoichiometry of Na+ to glutamate of 1:1, but further analysis showed that the stoichiometry cannot be resolved by purely kinetic studies. Studies with glutamate analogs, however, showed that kainic acid was a very effective stimulant of 22Na+ uptake, but 3H-kainic acid showed no Na+ -dependent uptake. Furthermore, while L-3H-glutamate uptake was very sensitive to lowered temperatures, glutamate-stimulated 22Na+ uptake was relatively insensitive. These results indicate that glutamate-stimulated uptake of 22Na+ in primary astrocytes cultures cannot be explained solely by cotransport of Na+ with glutamate, and they suggest that direct kainic acid-type receptor induced stimulation of Na+ uptake also occurs. Since both receptor and uptake effects involve transport of Na+, accurate measurements of the Na+ :glutamate stoichiometry for uptake can only be done using completely specific inhibitors of these 2 systems.

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The Journal of Neuroscience: 9 (4)
Journal of Neuroscience
Vol. 9, Issue 4
1 Apr 1989
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Excitatory amino acid-stimulated uptake of 22Na+ in primary astrocyte cultures
HK Kimelberg, S Pang, DH Treble
Journal of Neuroscience 1 April 1989, 9 (4) 1141-1149; DOI: 10.1523/JNEUROSCI.09-04-01141.1989

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Excitatory amino acid-stimulated uptake of 22Na+ in primary astrocyte cultures
HK Kimelberg, S Pang, DH Treble
Journal of Neuroscience 1 April 1989, 9 (4) 1141-1149; DOI: 10.1523/JNEUROSCI.09-04-01141.1989
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