Measurement of Glucose 6-Phosphate Dehydrogenase Activity in Bovine Oocytes and Embryos Using Qualitative Fluoresence Microscopy

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The activities of glucose 6-phosphate dehydrogenase (G6PDH) of the follicular oocytes collected from bovine ovaries and the embryos fertilized in vitro were measured using fluorescence microscope with photomultiplier and photometer attachments. The response for the fluorescence of NADPH was found to be linear, with correlation coefficiency of 0.997 and the limit of detection was 0.5mM.The G6PDH activity of intact oocytes could not be measured directly. Then the zona pellucida free follicular oocytes in extraction solution were frozen in LN_2, thawed and centrifuged and the G6PDH activity of their supernatant could be measured. Using this method, the G6PDH activities of oocytesat germinal vesicle and second metaphase stages and embryos at two-cell to morulae stages were determined. The G6PDH activities were 0.67 to 0.84 nmol NADPH generated/embryo/h at germinal vesicle stage to four-cell stage and the activity was decreased thereafter. There was no activity at morulae stage.