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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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