High developmental competence of pig oocytes after meiotic inhibition with a specific M-phase promoting factor kinase inhibitor, butyrolactone I
| Autor: | Guang-Ming, Wu, Qing-Yuan, Sun, Jiude, Mao, Liangxue, Lai, Tod C, McCauley, Kwang-Wook, Park, Randall S, Prather, Brad A, Didion, Billy N, Day |
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| Rok vydání: | 2002 |
| Předmět: |
Cell Nucleus
Cytoplasm Microscopy Confocal Swine Maturation-Promoting Factor Fertilization in Vitro Chromatin Mitochondria Enzyme Activation Actin Cytoskeleton Embryonic and Fetal Development Meiosis 4-Butyrolactone Oocytes Animals Female Enzyme Inhibitors Mitogen-Activated Protein Kinases Phosphorylation |
| Zdroj: | Biology of reproduction. 67(1) |
| ISSN: | 0006-3363 |
| Popis: | Butyrolactone I specifically inhibits M-phase promoting factor activation and prevents the resumption of meiosis. These experiments were conducted to examine effects of butyrolactone I on pig oocytes in a serum-free maturation system. The first experiment was conducted to determine the effect of butyrolactone I (0-100 microM) on nuclear maturation. At concentrations ofor =12.5 microM, germinal vesicle breakdown was prevented in90% of the oocytes after 24 h of culture. In the second experiment, the kinetics of in vitro maturation of butyrolactone I-treated oocytes was investigated. Oocytes were treated with 0 or 12.5 microM butyrolactone I and FSH for 20 h and then cultured with LH in the absence of butyrolactone I for another 24 h. Fewer butyrolactone I-treated oocytes reached MII stage at 36 h compared with controls (5.8% vs. 62.4%, P0.01). However, by 44 h, 83.4% of butyrolactone I-treated oocytes reached MII compared with 88.6% of controls. In the third experiment, butyrolactone I-treated oocytes were fertilized and cultured in vitro. No differences (P0.05) were found between controls and treated groups in cleavage rate, blastocyst rate, or mean number of cells per blastocyst. Effects of butyrolactone I on mitogen-activated protein kinase activation and localization of microfilaments and active mitochondria were examined by Western blot analysis and laser scanning confocal microscopy, respectively. The results suggested that although butyrolactone I reversibly inhibited germinal vesicle breakdown and mitogen-activated protein kinase activation, it did not affect mitochondrial and microfilament dynamics. Butyrolactone I is a potent inhibitor of nuclear maturation of porcine oocytes, and the inhibition is fully reversible. |
| Databáze: | OpenAIRE |
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