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Biology of Reproduction 61, 900-905 (1999)
©Copyright 1999 Society for the Study of Reproduction, Inc.

Articles

A Protein Tyrosine Phosphatase Inhibitor, Sodium Orthovanadate, Causes Parthenogenetic Activation of Pig Oocytes via an Increase in Protein Tyrosine Kinase Activity1

Jae-Hwan Kima, Hyun-Jin Doa, Wei-Hua Wang3,a, Zoltan Machátya, Yong-Mahn Han4,a, Billy N. Daya, and Randall S. Prather2,a

a Department of Animal Science, University of Missouri, Columbia, Missouri 65211

This study was conducted to determine whether a protein tyrosine kinase (PTK) activity is involved in the initiation of the events that occur at fertilization in pig oocytes. After maturation for 47 h, a 7-h treatment of oocytes with 1 mM sodium orthovanadate, which is an inhibitor of protein tyrosine phosphatase, caused more than 90% pronuclear formation, cortical granule exocytosis, and a decrease in mitogen-activated protein kinase activity. Immunoblotting with an antibody specific for phosphotyrosine showed at least three proteins whose phosphotyrosine contents were significantly increased upon treatment of oocytes with 1 mM sodium orthovanadate. Preincubation of pig oocytes with 50 µM tyrphostin 47, a specific PTK inhibitor, completely blocked the ability of sodium orthovanadate to trigger activation events. In addition, when oocytes were pretreated with the calcium-chelating agent BAPTA-AM, sodium orthovanadate-stimulated pronuclear formation was significantly (P < 0.01) reduced (94.0% vs. 43.1%). These results suggest that PTK may be involved in pig oocyte activation in a calcium-dependent manner and that the stimulation of tyrosine kinase is able to signal a series of intracellular changes that lead to the activation events associated with fertilization.

1 This material is based upon work supported by the Food for the 21st Century and the Cooperative State Research, Education and Extension Service, U.S. Department of Agriculture, under agreement No. 95-37203-2073. The manuscript is a contribution from the Missouri Agriculture Experiment Station Journal Series No. 12,918.

2 Correspondence: Randall S. Prather, 162 Animal Science Research Center, University of Missouri-Columbia, Columbia, MO 65211. FAX: 573 882 6827; pratherr{at}missouri.edu

3 Current address: IVF Lab., Women & Infant's Hospital of Rhode Island, Brown University School of Medicine, Providence, RI 02905.

4 Current address: Korean Research Institute of Bioscience & Biotechnology, Taejon 305-600, Korea.




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K.-i. Sato, T. Iwasaki, K. Ogawa, M. Konishi, A. A. Tokmakov, and Y. Fukami
Low density detergent-insoluble membrane of Xenopus eggs: subcellular microdomain for tyrosine kinase signaling in fertilization
Development, March 4, 2003; 129(4): 885 - 896.
[Abstract] [Full Text] [PDF]


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