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BOR - Papers in Press, published online ahead of print December 13, 2006.
Biol Reprod 2006, 10.1095/biolreprod.106.057828
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Submitted September 28, 2006
Returned for revision October 23, 2006
Accepted December 8, 2006

Gamete Biology


Adenosine 5’ Monophosphate Kinase-Activated Protein Kinase (PRKA) Activators Delay Meiotic Resumption in Porcine Oocytes

Mario A. Mayes , Martin F. Laforest , Christine Guillemette , Robert B. Gilchrist , and Francois J. Richard *

* To whom correspondence should be addressed. E-mail: francois.richard{at}crbr.ulaval.ca.

Abstract
Adenosine monophosphate-activated kinase (PRKA) is a serine/threonine kinase that functions as a metabolic switch in a number of physiological functions. This study was undertaken to assess the role of this kinase in nuclear maturation of porcine oocytes. We showed by RT-PCR and immunoblotting the expression of the PRKAA1 subunit in granulosa cells, COC and DO. Porcine COC and DO have transcripts corresponding to the expected size of the designed primers for PRKAB1 and PRKAG1. The PRKAA2 subunit was detected in granulosa cells and COC, whereas the PRKAG3 subunit was not detected in granulosa cells, COC and DO, but it was detected in heart. The PRKAA1 protein was detected in granulosa cells, COC, DO and zona pellucida (ZP). In the presence of the pharmacological activator of PRKA, ZMP (5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranosyl 5'-monophosphate), COC were transiently maintained in meiotic arrest in a fully reversible manner. This inhibitory effect was not observed in DO. Other known PRKA activators, AICAR (5-aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside) and metformin, also blocked meiotic resumption in COC. In contrast to mouse oocytes, where PRKA activators reverse the inhibitory effect of PDE3 inhibitors, this combination still blocked meiotic resumption in porcine COC. These results demonstrate that the meiotic resumption of porcine COC is transiently blocked by PRKA activators in a dose-dependant manner and that this effect is dependent on PRKA actions in cumulus cells. This study describes a new role for PRKA in regulating meiotic resumption in the COC and strongly suggests that cumulus cells play an essential role in the control of porcine oocyte maturation by the PRKA metabolic switch.

Key words: Gamete Biology • Follicle • Kinases • Meiosis • Ovum


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[Abstract] [Full Text] [PDF]


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