Pyruvate utilization by mouse oocytes is influenced by meiotic status and the cumulus oophorus.
In this study, the effects of meiotic status on the energy substrate dynamics of mouse oocyte-cumulus cell complexes (OCCs) and denuded oocytes (DOs) have been examined. In the first series of experiments, OCCs from PMSG-primed, immature mice were cultured in minimum essential medium in 8-microl microdrops under a variety of conditions, and the medium and oocytes were sampled for pyruvate and glucose concentration and for meiotic status. Oocytes in control medium underwent germinal vesicle breakdown within 3 hr and the OCCs displayed a time-dependent increase in pyruvate consumption, but the glucose concentration changed very little. Treatment with IBMX or dbcAMP, which maintained complete meiotic arrest, suppressed pyruvate consumption, but slightly more glucose was consumed than in controls. Hypoxanthine (HX) allowed up to 10% of the oocytes to resume maturation, and pyruvate and glucose consumption resembled that of control OCCs. FSH added to HX-containing medium stimulated significant glucose consumption and pyruvate production. In general, a reciprocal relationship was observed between glucose and pyruvate consumption. When the energy substrate dynamics were compared with meiotic status of the oocytes, pyruvate consumption was associated with the maturation process. Although HX maintained oocytes in the germinal vesicle stage, the meiotic arrest was "leaky," allowing increased pyruvate consumption. Additional experiments showed that DOs at either the prophase I or metaphase II stages consumed less pyruvate than oocytes actively engaged in meiotic maturation. DOs oxidized significantly more pyruvate than OCCs, and glycolytic metabolism of glucose lowered the oxidation rate in OCCs. Furthermore, while 5-6.2 times more pyruvate was consumed by OCCs than by DOs in the absence of glucose, oxidation did not mediate the meiosis-inducing effect of pyruvate, since less of this substrate was oxidized by OCCs than by DOs. We conclude that meiotically active oocytes have a greater requirement for pyruvate than prophase I- or metaphase II-arrested oocytes and that meiotic status can influence the metabolism not only of oocytes, but also of the OCCs.
Downs
Stephen M
SM
Biology Department, Marquette University, 530 N 15 Street, Milwaukee, WI 53233, USA. stephen.downs@marquette.edu
Humpherson
Peter G
PG
Leese
Henry J
HJ
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D000818:Animals; D003994:Bucladesine / Q000494:pharmacology; D002454:Cell Differentiation; D002478:Cells, Cultured; D005260:Female; D019271:Hypoxanthine / Q000494:pharmacology; D008540:Meiosis / Q000502:physiology*; D051379:Mice; D008810:Mice, Inbred C57BL; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D010053:Ovary; D010084:Oxidation-Reduction; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D019289:Pyruvic Acid / Q000378:metabolism*
D010726:Phosphodiesterase Inhibitors; D019271:Hypoxanthine; D003994:Bucladesine; D019289:Pyruvic Acid; D015056:1-Methyl-3-isobutylxanthine