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Proteomic analysis of porcine oocytes during in vitro maturation reveals essential role for the ubiquitin C-terminal hydrolase-L1.
- Source :
-
Reproduction (Cambridge, England) [Reproduction] 2007 Oct; Vol. 134 (4), pp. 559-68. - Publication Year :
- 2007
-
Abstract
- In this study, we performed proteomic analysis of porcine oocytes during in vitro maturation. Comparison of oocytes at the initial and final stages of meiotic division characterized candidate proteins that were differentially synthesized during in vitro maturation. While the biosynthesis of many of these proteins was significantly decreased, we found four proteins with increased biosynthetic rate, which are supposed to play an essential role in meiosis. Among them, the ubiquitin C-terminal hydrolase-L1 (UCH-L1) was identified by mass spectrometry. To study the regulatory role of UCH-L1 in the process of meiosis in pig model, we used a specific inhibitor of this enzyme, marked C30, belonging to the class of isatin O-acyl oximes. When germinal vesicle (GV) stage cumulus-enclosed oocytes were treated with C30, GV breakdown was inhibited after 28 h of culture, and most of the oocytes were arrested at the first meiosis after 44 h. The block of metaphase I-anaphase transition was not completely reversible. In addition, the inhibition of UCH-L1 resulted in elevated histone H1 kinase activity, corresponding to cyclin-dependent kinase(CDK1)-cyclin B1 complex, and a low level of monoubiquitin. These results supported the hypothesis that UCH-L1 might play a role in metaphase I-anaphase transition by regulating ubiquitin-dependent proteasome mechanisms. In summary, a proteomic approach coupled with protein verification study revealed an essential role of UCH-L1 in the completion of the first meiosis and its transition to anaphase.
- Subjects :
- Animals
CDC2 Protein Kinase analysis
CDC2 Protein Kinase metabolism
Egg Proteins biosynthesis
Electrophoresis, Gel, Two-Dimensional
Female
Fertilization in Vitro
Immunoblotting
Indoles pharmacology
Meiosis drug effects
Myelin Basic Protein analysis
Myelin Basic Protein metabolism
Oocytes metabolism
Oximes pharmacology
Proteasome Endopeptidase Complex
Protein Kinases analysis
Protein Kinases metabolism
Spectrum Analysis
Swine
Ubiquitin metabolism
Ubiquitin Thiolesterase analysis
Ubiquitin Thiolesterase antagonists & inhibitors
Egg Proteins analysis
Meiosis physiology
Oocytes chemistry
Ubiquitin Thiolesterase metabolism
Subjects
Details
- Language :
- English
- ISSN :
- 1470-1626
- Volume :
- 134
- Issue :
- 4
- Database :
- MEDLINE
- Journal :
- Reproduction (Cambridge, England)
- Publication Type :
- Academic Journal
- Accession number :
- 17890291
- Full Text :
- https://doi.org/10.1530/REP-07-0079