Butyrolactone I reversibly inhibits meiotic maturation of bovine oocytes,Without influencing chromosome condensation activity
Language English Country United States Media print
Document type Journal Article, Research Support, Non-U.S. Gov't, Research Support, U.S. Gov't, P.H.S.
Grant support
HD22681
NICHD NIH HHS - United States
- MeSH
- Chromatin ultrastructure MeSH
- Chromosomes drug effects MeSH
- Electrophoresis, Polyacrylamide Gel MeSH
- Myelin Basic Protein metabolism MeSH
- Fertilization physiology MeSH
- Tissue Fixation MeSH
- Follicular Fluid cytology MeSH
- 4-Butyrolactone analogs & derivatives pharmacology MeSH
- Enzyme Inhibitors pharmacology MeSH
- Culture Media MeSH
- Cells, Cultured MeSH
- Meiosis drug effects MeSH
- Mitogen-Activated Protein Kinases antagonists & inhibitors metabolism MeSH
- Oocytes drug effects enzymology growth & development MeSH
- CDC2 Protein Kinase metabolism MeSH
- Protein Kinases metabolism MeSH
- Cattle MeSH
- Spermatozoa physiology MeSH
- Blotting, Western MeSH
- Germinal Center drug effects MeSH
- Animals MeSH
- Check Tag
- Male MeSH
- Cattle MeSH
- Female MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Research Support, U.S. Gov't, P.H.S. MeSH
- Names of Substances
- butyrolactone I MeSH Browser
- Chromatin MeSH
- Myelin Basic Protein MeSH
- 4-Butyrolactone MeSH
- histone H1 kinase MeSH Browser
- Enzyme Inhibitors MeSH
- Culture Media MeSH
- Mitogen-Activated Protein Kinases MeSH
- CDC2 Protein Kinase MeSH
- Protein Kinases MeSH
In this study, butyrolactone I (BL I), a potent and specific inhibitor of cyclin-dependent kinases, was shown to block germinal vesicle (GV) breakdown (GVBD) in bovine oocytes in a concentration-dependent manner; GVBD was almost totally inhibited over the course of 24-48 h of culture when 100 microM BL I was included in tissue culture medium 199 containing either polyvinyl alcohol or BSA. Correlated with this inhibition was the failure of either p34(cdc2) kinase or mitogen-activated protein (MAP) kinase to become activated, and it was unlikely that BL I directly inhibited MAP kinase, since 100 microM BL I did not inhibit MAP kinase activity present in extracts obtained from metaphase II-arrested bovine eggs that possess high levels of MAP kinase activity. Nevertheless, the formation of highly condensed bivalents was observed in 78% of the BL I-treated GV-intact oocytes. This result suggests that chromosome condensation during first meiosis in bovine oocytes does not require the activity of either p34(cdc2) kinase or MAP kinase. Treatment of BL I-arrested oocytes with okadaic acid (OA) did not result in either the activation of p34(cdc2) kinase or MAP kinase, or inducement of GVBD. The BL I-induced block of GVBD for 24 h was reversible, and a subsequent 24-h culture resulted in 90% of oocytes reaching metaphase II with emission of the first polar body. Correlated with the progression to and arrest at metaphase II was the full activation of both p34(cdc2) and MAP kinases. The reversibility after 48 h of culture in BL I was partially decreased when compared to that achieved after an initial 24-h culture. Fertilization in vitro of these eggs resulted in a high incidence of both sperm penetration and pronucleus formation (88% and 70%, respectively).
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