Membrane Estrogen Receptor-α Interacts with Metabotropic Glutamate Receptor Type 1a to Mobilize Intracellular Calcium in Hypothalamic Astrocytes

Estradiol, acting on a membrane-associated estrogen receptor-alpha (mER alpha), induces an increase in free cytoplasmic calcium concentration ([Ca2+](i)) needed for progesterone synthesis in hypothalamic astrocytes. To determine whether rapid estradiol signaling involves an interaction of mER alpha with metabotropic glutamate receptor type 1a (mGluR1a), changes in [Ca2+](i) were monitored with the calcium indicator, Fluo-4 AM, in primary cultures of female postpubertal hypothalamic astrocytes. 17 beta-Estradiol over a range of 1 nM to 100 nM induced a maximal increase in [Ca2+](i) flux measured as a change in relative fluorescence [Delta F Ca2+ = 615 +/- 36 to 641 +/- 47 relative fluorescent units (RFU)], whereas 0.1 nM of estradiol stimulated a moderate [Ca2+](i) increase (275 +/- 16 RFU). The rapid estradiol-induced [Ca2+](i) flux was blocked with 1 mu M of the estrogen receptor antagonist ICI 182,780 (635 +/- 24 vs. 102 +/- 11 RFU, P < 0.001) and 20 nM of the mGluR1a antagonist LY 367385 (617 +/- 35 vs. 133 +/- 20 RFU, P < 0.001). Whereas the mGluR1a receptor agonist (RS)-3,5-dihydroxyphenyl-glycine (50 mu M) also stimulated a robust [Ca2+](i) flux (626 +/- 23 RFU), combined treatment of estradiol (1 nM) plus (RS)-3,5-dihydroxyphenyl-glycine (50 mu M) augmented the [Ca2+](i) response (762 +/- 17 RFU) compared with either compound alone (P < 0.001). Coimmunoprecipitation demonstrated a direct physical interaction between mER alpha and mGluR1a in the plasma membrane of hypothalamic astrocytes. These results indicate that mER alpha acts through mGluR1a, and mGluR1a activation facilitates the estradiol response, suggesting that neural activity can modify estradiol-induced membrane signaling in astrocytes. (Endocrinology 150: 1369-1376, 2009)