Chemical manipulation of glucose metabolism in porcine oocytes: effects on nuclear and cytoplasmic maturation in vitro.
The objectives of this study were to manipulate metabolism of glucose through glycolysis and the pentose phosphate pathway (PPP) in porcine oocytes during in vitro maturation, and determine the effects of this manipulation on meiotic progression, intracellular glutathione (GSX) concentrations and embryonic development. Cumulus-oocyte complexes isolated from abattoir ovaries were matured (40-44 h) in Purdue Porcine Medium for maturation alone (control) or supplemented with pyrroline-5 carboxylate (PC, 0.1 microM; PPP stimulator), diphenyleneiodonium (DPI, 0.1 microM; PPP inhibitor), dinitrophenol (DNP, 10 microM; glycolytic stimulator), hexametaphosphate (HMP, 100 microM; glycolytic inhibitor), PC + HMP or DNP + DPI. At the conclusion of in vitro maturation, cumulus cells were removed and oocytes were randomly allocated for analysis of GSX, metabolism and nuclear maturation, or in vitro fertilization and embryo culture. Both DPI and DNP + DPI decreased (P < or = 0.05) the activity of glycolysis and the PPP, increased (P < or = 0.05) the percentage of immature oocytes, and decreased (P < or = 0.05) the proportion of mature oocytes compared with control oocytes and oocytes from the other treatments. Embryonic development (cleavage and blastocyst stage) and the intracellular content of GSX were also decreased (P < or = 0.05) following exposure to DPI or DNP + DPI compared with control oocytes and oocytes from the other treatments. Oocyte metabolism, nuclear maturation, GSX content and embryonic development were unaffected (P > 0.05) following exposure to PC, DNP, HMP or PC + HMP. Our results suggest that metabolism of glucose through the PPP and/or glycolysis plays a key role in the control of nuclear and cytoplasmic maturation of porcine oocytes in vitro.
Herrick
Jason R
JR
Department of Animal Sciences, Program of Comparative Medicine, Lilly Hall of Life Sciences,Purdue University, West Lafayette, Indiana 47907, USA.
Brad
Amber M
AM
Krisher
Rebecca L
RL
D000818:Animals; D002467:Cell Nucleus / Q000502:physiology*; D002478:Cells, Cultured; D003593:Cytoplasm / Q000502:physiology*; D004140:Dinitrophenols / Q000494:pharmacology; D047108:Embryonic Development; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods; D005947:Glucose / Q000378:metabolism*; D005978:Glutathione / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects; D042541:Intracellular Space / Q000378:metabolism; D008540:Meiosis / Q000187:drug effects; D009861:Onium Compounds / Q000494:pharmacology; D009865:Oocytes / Q000378:metabolism*; D009866:Oogenesis / Q000502:physiology*; D010427:Pentose Phosphate Pathway / Q000187:drug effects; D010710:Phosphates / Q000494:pharmacology; D013552:Swine / Q000502:physiology*; D014475:Uncoupling Agents / Q000494:pharmacology
D004140:Dinitrophenols; D004791:Enzyme Inhibitors; D009861:Onium Compounds; D010710:Phosphates; D014475:Uncoupling Agents; C005035:hexametaphosphate; C007517:diphenyleneiodonium; D005978:Glutathione; D005947:Glucose