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First published on August 30, 2005; DOI: 10.1124/mol.104.010124

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Received for publication January 10, 2005.
Revised July 26, 2005.
Accepted for publication August 30, 2005.

Intracellular cAMP and calcium signaling by serotonin in mouse cumulus-oocyte complexes

Pascal Amireault 1 Francois Dube 1*

1 University of Montreal

* Address correspondence to: E-mail: francois.dube{at}umontreal.ca

Abstract

Cyclic AMP (cAMP) and intracellular Ca2+ are important second messengers involved in mammalian follicular growth and oocyte meiotic maturation. We investigated the capacity of the neurohormone serotonin(or 5-hydroxytryptamine, 5-HT) to regulate intracellular cAMP and Ca2+ in mouse oocytes and surrounding cumulus cells. Based on a RT-PCR study, 5-HT7 receptor mRNA is expressed in cumulus cells, oocytes and embryos up to the 4 cell-stage and 5-HT2A and 5-HT2B receptor mRNAs in cumulus cells only, while 5-HT2C, 5-HT4 and 5-HT6 receptors are expressed neither in oocytes nor cumulus cells. The addition of 5-HT (10 nM-10 µM) to isolated metaphase II-oocytes had no effect on their internal cAMP or Ca2+ levels whereas it caused dose-dependent cAMP and Ca2+ increases in cumulus cells. This cAMP increase in cumulus cells could be mimicked by 5-HT agonists with the following order of potency: 5-HT > 8-OH DPAT = {alpha}-methyl 5-HT = 5-CT, thereby supporting a preferential involvement of 5-HT7 receptors. As measured with cumulus cells pre-loaded with fura2-AM, the addition of 5-HT also caused dose-dependent Ca2+ rises, likely linked to detected 5-HT2A and 5-HT2B receptors. Adding the Ca2+ ionophore ionomycin to cumulus cells resulted in both Ca2+ and cAMP rises whereas preincubation of cells with the Ca2+ chelator BAPTA-AM abolished the 5-HT induced Ca2+ rise and reduced the cAMP increase, thus indicating a cross-talk between the 5-HT-sensitive Ca2+ and cAMP pathways. Our results show that 5-HT may be a local regulator in mouse cumulus-oocyte complexes through its actions on cAMP and Ca2+ signaling, as mediated by 5-HT2A, 5-HT2B and 5-HT7 receptors.


Key words: Serotonin, cAMP, Calcium (G Protein Coupled Signals)


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[Abstract] [Full Text] [PDF]


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