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BOR - Papers in Press, published online ahead of print February 21, 2007.
Biol Reprod 2007, 10.1095/biolreprod.106.059899
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Submitted December 30, 2006
Returned for revision January 15, 2007
Accepted February 15, 2007

Gamete Biology


Oxidative Metabolism of Pyruvate Is Required for Meiotic Maturation of Murine Oocytes In Vivo

Mark T. Johnson *, Edward A. Freeman , David K. Gardner , and Patricia A. Hunt

* To whom correspondence should be addressed. E-mail: mtj{at}cwru.edu.

Abstract
The requirement for oxidative metabolism of pyruvate during oogenesis in vivo was evaluated by inactivating Pdha1, a gene encoding an enzymatic subunit of pyruvate dehydrogenase complex, in murine oocytes at the beginning of the follicular growth phase. Immunohistochemical analysis revealed that Pdha1- oocytes have dramatically reduced amounts of pyruvate dehydrogenase enzyme by the secondary follicle stage. Despite this reduction, these oocytes grow to normal size, are ovulated and can be fertilized. Pdha1- oocytes are, however, impaired in their ability to support embryonic development as demonstrated by the failure of fertilized oocytes to develop beyond the 1-cell zygote stage in vivo. Immunocytochemical evaluation showed that almost all (98.4%) ovulated Pdha1- oocytes have not completed meiotic maturation and/or have gross abnormalities of the meiotic spindle and chromatin. Meiotic maturation is even more compromised when these oocytes are matured in vitro in the absence of cumulus cells or in the presence of the gap junction inhibitor 18-alpha glycyrrhetinic acid, indicating that cumulus cells can partially compensate for this enzymatic deficiency through a gap junction-mediated mechanism. Ovulated Pdha1- oocytes were also shown to have reduced levels of total ATP content and NAD(P)H autofluorescence relative to oocytes without this enzymatic deficiency. These studies demonstrate that oxidative metabolism of pyruvate is essential for proper completion of oogenesis, serving as a vital source of energy during meiotic maturation. At earlier stages of oogenesis, this metabolic pathway may not be necessary due to metabolic compensation by the granulosa cells.

Key words: Embryo • Gamete Biology • Granulosa cells • Meiosis • Oocyte development


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[Abstract] [Full Text] [PDF]




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