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Fluorescence measurement of changes in intracellular calcium induced by excitatory amino acids in cultured cortical astrocytes

AM Jensen and SY Chiu
Journal of Neuroscience 1 April 1990, 10 (4) 1165-1175; DOI: https://doi.org/10.1523/JNEUROSCI.10-04-01165.1990
AM Jensen
Department of Neurophysiology, University of Wisconsin, Madison 53706.
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SY Chiu
Department of Neurophysiology, University of Wisconsin, Madison 53706.
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Abstract

Population response of [Ca2+]i in cultured cortical astrocytes to excitatory amino acids was measured at room temperature using the calcium-sensitive dye fura-2. Quisqualic acid (QA), glutamate (Glu), and kainic acid (KA) caused a peak increase in [Ca2+]i in the order QA greater than Glu greater than KA. No response to N-methyl-D-aspartic acid (NMDA) was observed whether or not Mg2+ was present externally. Both QA and Glu (100 microM) frequently elicited a decaying oscillatory [Ca2+]i response during sustained agonist application; the period of oscillations initially was 23.5 sec and increased as the response was damped. Comparatively, the [Ca2+]i response to KA was nonoscillatory. Both responses to Glu and KA were reduced slightly by antagonist gamma- D-glutamylaminomethyl-sulfonic acid (1 mM), but virtually were abolished by kynurenic acid (3 mM). Replacement of external Na+ by choline had no significant effect on the Glu response. Removal of external Ca2+ reduced the peak response to QA, Glu, and KA to 40, 34, and 18%, respectively; and markedly reduced the degree of QA- and Glu- induced [Ca2+]i oscillations. Pretreatment with phorbol esters, a potent activator of protein kinase C, blocked the [Ca2+]i response to Glu but not KA. It is concluded that cortical astrocytes express Glu receptors of the non-NMDA type in culture and that receptor activation leads to Ca2+ influx and release of internal Ca2+. Mobilization of Ca2+ apparently occurs via the known Glu-mediated hydrolysis of inositol lipids, which may come under negative-feed-back control by protein kinase C activation. Oscillatory [Ca2+]i signaling offers the possibility of a dynamic population response in an electrically coupled glial network.

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The Journal of Neuroscience: 10 (4)
Journal of Neuroscience
Vol. 10, Issue 4
1 Apr 1990
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Fluorescence measurement of changes in intracellular calcium induced by excitatory amino acids in cultured cortical astrocytes
AM Jensen, SY Chiu
Journal of Neuroscience 1 April 1990, 10 (4) 1165-1175; DOI: 10.1523/JNEUROSCI.10-04-01165.1990

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Fluorescence measurement of changes in intracellular calcium induced by excitatory amino acids in cultured cortical astrocytes
AM Jensen, SY Chiu
Journal of Neuroscience 1 April 1990, 10 (4) 1165-1175; DOI: 10.1523/JNEUROSCI.10-04-01165.1990
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