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Reversible changes in protein phosphorylation during germinal vesicle breakdown and pronuclear formation in bovine oocytes in vitro

Published online by Cambridge University Press:  06 June 2003

R.C. Chian
Affiliation:
Department of Obstetrics and Gynecology, McGill University, Montreal, Canada H3A 1A1 Department of Animal Science, Macdonald Campus, McGill University, Ste-Anne de Bellevue, Canada H9X 3V9
J.T. Chung
Affiliation:
Department of Obstetrics and Gynecology, McGill University, Montreal, Canada H3A 1A1
K. Niwa
Affiliation:
Division of Animal Science and Technology, Faculty of Agriculture, Okayama University, 700-8530 Okayama, Japan
M.A. Sirard
Affiliation:
Department of Animal Science, University of Laval, Quebec, Canada G1K 7P4
B.R. Downey
Affiliation:
Department of Animal Science, Macdonald Campus, McGill University, Ste-Anne de Bellevue, Canada H9X 3V9
S.L. Tan
Affiliation:
Department of Obstetrics and Gynecology, McGill University, Montreal, Canada H3A 1A1

Abstract

This study examined the event of protein phosphorylation in bovine oocytes during germinal vesicle breakdown (GVBD) and formation of pronuclei following fertilisation in vitro. Immature oocytes were obtained from abattoir materials and cultured in vitro. The oocytes were labelled with [32P]orthophosphate at 3 h intervals from 0 to 12 h following maturation in culture or from 3 to 18 h following insemination. One-dimensional gel electrophoresis indicated that levels of protein phosphorylation are low prior to GVBD. However, the levels of protein phosphorylation at approximately 40 kDa, 27 kDa, 23 kDa and 18 kDa increased substantially following GVBD and then decreased gradually as maturation in culture progressed. In contrast, the levels of protein phosphorylation increased gradually in the oocytes following pronucleus formation. Further, two-dimensional gel electrophoresis indicated that the protein at approximately 18 kDa reversibly changed in the oocytes during maturation and fertilisation. These results indicate that the reversible changes of this phosphoprotein may be related to either cell cycle transition or pronucleus formation during maturation and fertilisation in bovine oocytes.

Type
Research Article
Copyright
2003 Cambridge University Press

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