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A Membrane Estrogen Receptor Mediates Intracellular Calcium Release in Astrocytes

Laboratory of Neuroendocrinology of the Brain Research Institute, Department of Neurobiology, Mental Retardation Research Center, and Center for Neurovisceral Sciences and Woman’s Health, David Geffen School of Medicine at University of California, Los Angeles, Los Angeles, California 90095-1763

Address all correspondence and requests for reprints to: Victor V. Chaban, Department of Neurobiology, David Geffen School of Medicine at University of California, Los Angeles, 10833 Le Conte Avenue, Los Angeles, California 90095-1763. E-mail: chaban{at}ucla.edu.

Appreciating the physiology of astrocytes and their role in brain functions requires an understanding of molecules that activate these cells. Estradiol may influence astrocyte functions. We now report that estrogen altered intracellular calcium concentration ([Ca²⁺]_i) in neonatal astrocytes that expressed estrogen receptor (ER) mRNA in vitro. Western blotting revealed both ER and ERß proteins in both the nuclear fractions and plasma-membrane fractions. Application of 17ß-estradiol (20 nM) to fura 2-loaded astrocytes in vitro stimulated [Ca²⁺]_i in 75% of astrocytes with an EC₅₀ of 12.7 ± 3.1 nM. This rapid action of estradiol was blocked by the ER antagonist, ICI 182,780. The membrane-impermeable estradiol-BSA induced a [Ca²⁺]_i flux that was statistically similar to estradiol. Removal of extracellular Ca²⁺ did not alter the effect of estradiol, but phospholipase C inhibitor U73122 (10 µM) and 2-aminoethoxydiphenyl borate (5 µM), an inhibitor of the inositol-1,4,5,-trisphosphate-gated intracellular Ca²⁺ channel, significantly decreased the estradiol-induced [Ca²⁺]_i flux. Estradiol was unable to induce [Ca²⁺]_i flux in thapsigargin-depleted cells. These results indicate that estradiol mediates [Ca²⁺]_i flux in astrocytes through a membrane-associated ER that activates the phospholipase C pathway.

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