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Hum. Reprod. Advance Access published online on June 2, 2005
Human Reproduction, doi:10.1093/humrep/dei078
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© The Author 2005. Published by Oxford University Press on behalf of the European Society of Human Reproduction and Embryology. All rights reserved. For Permissions, please email: journals.permissions@oupjournals.org
Received October 27, 2004
Revised March 18, 2005
Accepted April 14, 2005

Non-genomic action of estradiol and progesterone on cytosolic calcium concentrations in primary cultures of human granulosa-lutein cells

E.V. Younglai 1*, Y.J. Wu 1, T.K. Kwan 2, and C.-Y. Kwan 2

1 Department of Obstetrics and Gynecology, Reproductive Biology Division, McMaster University, Health Sciences Centre, Hamilton, Ontario, Canada L8N 3Z5
2 Department of Medicine, McMaster University, Health Sciences Centre, Hamilton, Ontario, Canada L8N 3Z5

* To whom correspondence should be addressed.
E.V. Younglai, E-mail: younglai{at}mcmaster.ca


  Abstract

BACKGROUND: The present study examined whether the sex steroids, estradiol and progesterone, could alter cytoplasmic calcium concentrations ([Ca2+]cyt) in human granulosa-lutein cells. METHODS: Human granulosa cells were obtained at the time of oocyte retrieval for IVF and cultured for 3-7 days. Cells were loaded with Fura-2 AM and changes in [Ca2+]cyt of single cells were studied using a dynamic digital Ca2+ imaging system. RESULTS: Both estradiol and progesterone stimulated elevations of [Ca2+]cyt in Ca2+-containing medium within seconds of exposure of the granulosa-lutein cells to the steroid, but only estradiol caused an increase in [Ca2+]cyt in Ca2+-free medium. Both ICI-182780 and RU 486 stimulated [Ca2+]cyt increases and inhibited the effects of estradiol and progesterone, respectively. Tamoxifen also induced transient increases in [Ca2+]cyt concentrations but inhibited the effects of both estradiol and progesterone. The inhibitory effects of tamoxifen, ICI-182780 and RU 4486 on [Ca2+]cyt responses to estradiol and progesterone could be reversed with higher concentrations of estradiol and progesterone, respectively. The [Ca2+]cyt effects induced with tamoxifen could not be eliminated by prior treatment with RU 486 or ICI-182780. CONCLUSION: These results provide strong evidence that both estradiol and progesterone as well as the steroid antagonists, tamoxifen, RU 486 and ICI-182780, can act on human granulosa-lutein cells through a non-genomic mechanism.

Keywords: calcium response; estradiol; granulosa cells; progesterone.
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