Carbohydrate metabolism by murine ovarian follicles and oocytes grown in vitro.
Metabolic markers are potentially valuable for assessment of follicle development in vitro. Carbohydrate metabolism of murine preantral follicles grown to maturity over 13 days in vitro has been measured, and metabolism of resulting oocyte-cumulus complexes (OCCs) and denuded oocytes has been compared with in vivo ovulated control counterparts. Spent follicle culture media were analysed for glucose, lactate and pyruvate concentrations. During follicle in vitro growth, glycolysis accounted for a rise from approximately 24 to 60% of all glucose consumed. Ovulation induction caused a significant increase in glucose uptake and lactate production by in vitro-grown follicles to 71.7+/-1.2 and 96.6+/-4.8 nmoles/day respectively. OCCs grown in vitro had significantly higher rates of glucose consumption and lactate and pyruvate production (110.1+/- 3.5, 191.8+/- 8.9 and 31.7+/- 1.7 pmoles/h respectively) than in vivo ovulated controls (67.4+/- 8.1, 113.9+/- 17.1 and 20.2+/- 4.0 pmoles/h respectively), but a reduced capacity for pyruvate consumption (1.13+/- 0.06 vs 1.49+/- 0.06 pmoles/h by in vivo ovulated oocytes). Metabolism of OCCs was affected by the quality of the original follicle. In vitro-grown oocytes had a reduced cytoplasmic volume when compared with controls (168.3+/- 2.0 vs 199.0+/- 3.2 proportionately respectively) but a similar rate of metabolism per unit volume. Meiotic status influenced metabolism of both OCCs and denuded oocytes. In conclusion, glucose consumption and lactate production by cultured follicles increased in tandem with developmental progression and were stimulated prior to ovulation. Additionally, the metabolic profiles of in vitro produced OCCs and the oocytes within them are affected by long-term exposure to the culture environment.
Harris
Sarah E
SE
Reproduction and Early Development Research Group, Leeds Institute of Genetics, Health and Therapeutics, University of Leeds, Leeds LS2 9JT, UK. s.e.harris@leeds.ac.uk
Adriaens
Iris
I
Leese
Henry J
HJ
Gosden
Roger G
RG
Picton
Helen M
HM
D000818:Animals; D015415:Biomarkers / Q000032:analysis; D050260:Carbohydrate Metabolism; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000737:chemistry; D003600:Cytosol / Q000648:ultrastructure; D005260:Female; D005947:Glucose / Q000032:analysis; D019344:Lactic Acid / Q000032:analysis; D008540:Meiosis / Q000502:physiology; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000378:metabolism* / Q000648:ultrastructure; D006080:Ovarian Follicle / Q000378:metabolism*; D010062:Ovulation Induction; D011773:Pyruvates / Q000032:analysis
D015415:Biomarkers; D017077:Culture Media, Conditioned; D011773:Pyruvates; D019344:Lactic Acid; D005947:Glucose