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The Journal of Clinical Endocrinology & Metabolism Vol. 83, No. 1 263-269
Copyright © 1998 by The Endocrine Society

Original Studies

Nongenomic Effects of Androstenedione on Human Granulosa Luteinizing Cells

Véronique Machelon, Françoise Nomé and Jan Tesarik

INSERM, U-355, Institut Paris-Sud sur les Cytokines (V.M., F.N.), 92140 Clamart; Laboratoire d’Eylau (J.T.), 75116 Paris, France

Address all correspondence and requests for reprints to: Dr. Véronique Machelon, INSERM, U-355, Maturation Gamétique et Fécondation, 32 rue des Carnets, 92140 Clamart, France.

This study examines rapid (5–60 s) effects of androgens on the cytosolic free Ca2+ concentration ([Ca2+]i) in human granulosa luteinizing cells. Cells were obtained from human preovulatory follicles, and [Ca2+]i was measured with the use of the Ca2+-responsive fluorescent dye fluo-3. Molar concentrations between 100 pmol/L and 1 µmol/L androstenedione increased [Ca2+]i within 5 s after addition to cells. This [Ca2+]i increase resulted from both Ca2+ influx, as shown by the effects of ethyleneglycol-bis-(ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid and the voltage-dependent Ca2+ channel blocker verapamil, and Ca2+ mobilization from the endoplasmic reticulum, as shown by the effects of thapsigargin. Treatment with pertussis toxin and U-73,122, a specific inhibitor of phospholipase C, abolished the effects of androstenedione on [Ca2+]i. Flutamide, a nuclear androgen receptor antagonist, did not block the increase in [Ca2+]i induced by androstenedione. Testosterone (100 pmol/L to 1 µmol/L) had no effect. This is the first report showing that androstenedione increases [Ca2+]i in granulosa cells. These data provide evidence for the presence in granulosa cells of a novel, short term mechanism of androstenedione action involving voltage-dependent Ca2+ channels in the plasma membrane and phospholipase C activation via a pertussis toxin-sensitive G protein.




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