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Journal of Neuroscience, Vol 10, 1165-1175, Copyright © 1990 by Society for Neuroscience
Fluorescence measurement of changes in intracellular calcium induced by excitatory amino acids in cultured cortical astrocytes
AM Jensen and SY Chiu
Department of Neurophysiology, University of Wisconsin, Madison 53706.
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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