Differential effects of kinase inhibitor and electrical stimulus on activation and histone H1 kinase activity in pig oocytes.
The 6-DMAP (6-dimethylaminopurine) has been reported to accelerate and enhance the formation of pronuclei (activation) in non-aged MII mouse and bovine eggs. In this study, effects of chemical activation (CA) were evaluated using ethanol + 6-DMAP and electrical activation (EA) on pig oocytes matured for 36 h (newly matured) and 48 h. After CA, the first pronuclei developed within 2 h, with maximal pronuclear formation at 8 h (> 90%) for oocytes matured for either 36 or 48 h. In addition, chemically activated oocytes did not extrude the second polar body. After EA treatment, pronuclear formation began at 4-6 h and was significantly lower at 8 h in newly matured than aged oocytes (34% vs. 85%). Cleavage rate at 24 h after treatment was lower for oocytes treated by CA (< 50%) than for EA (> 70%) and after 3 days of in vitro culture, fewer oocytes reached the four-cell stage in CA than in EA groups. No chemically activated oocytes developed beyond the four-cell stage; in contrast, EA resulted in development beyond this stage. Both CA and EA resulted in a drop of H1 kinase activity, but EA induced a more rapid reduction. With EA, H1 kinase activity rapidly declined but tended to increase again at 8 h in oocytes matured for 36 h, which was different from all other groups in which the kinase activity remained low at 8 h post-activation. We conclude that different mechanisms are involved in pronuclear formation with CA and EA and that 6-DMAP-activated oocytes may not be suitable for the induction of normal parthenogenetic development in pigs. It is also suggested that the ability of pig oocytes to become fully activated may be related to the modification of H1 kinase activity during the aging process. These modifications to H1 kinase may be necessary for parthenogenetic activation of in vitro matured oocytes and also for the completion of meiosis and cleavage.
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Department of Development and Signaling, The Babraham Institute, Cambridge, UK.
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D000225:Adenine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D004558:Electric Stimulation; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D000431:Ethanol / Q000494:pharmacology; D005260:Female; D008856:Microscopy, Fluorescence / Q000662:veterinary; D008858:Microscopy, Phase-Contrast / Q000662:veterinary; D009865:Oocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D010312:Parthenogenesis / Q000187:drug effects; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000032:analysis / Q000378:metabolism*; D013552:Swine / Q000502:physiology*
D004791:Enzyme Inhibitors; D047428:Protein Kinase Inhibitors; D000431:Ethanol; C001020:N(6),N(6)-dimethyladenine; D011494:Protein Kinases; C102333:histone H1 kinase; D000225:Adenine