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Connexin43 in Rat Oocytes: Developmental Modulation of Its Phosphorylation¹

^a In Vitro Fertilization Unit, Obstetric and Gynecology Department, Kaplan Medical Center, 76100 Rehovot, Israel ^b Department of Biological Regulation, The Weizmann Institute of Science, 76100 Rehovot, Israel

It is well established that the 43-kDa connexin (Cx43) is predominantly expressed by ovarian somatic cells, whereas the identity of the connexins contributed by the oocyte to form gap junctions with its neighboring cells is not fully elucidated. Our study aimed to examine oocytes for the expression and regulation of Cx43 throughout oogenesis. Growing and fully grown rat oocytes that were meiotically incompetent and competent, respectively, were examined. Fully grown oocytes were analyzed either before or after reinitiation of meiosis as well as at the second meiotic metaphase. Immunofluorescent analysis of zona pellucida-free oocytes using conventional and confocal microscopy demonstrated a characteristic pattern of punctuated staining of Cx43 on the oolema. Immunogold electron microscopy localized Cx43 to the oocyte surface and the microvillar processes. Reverse transcriptase-polymerase chain reaction and Western blot analysis revealed similar amounts of Cx43 gene and protein in oocytes of different developmental stages. However, a relative increase in the phosphorylated forms of the protein was observed in fully grown oocytes that had completed their maturation. Our findings demonstrate that rat oocytes express a developmentally regulated Cx43. They further suggest that homotypic gap junctions that consist of Cx43 may be present between rat oocytes and their adjacent cumulus cells.

First decision: 6 July 2001.

¹ This work was partially supported by grants from the Maria Zondek Hormone Research Fund, the Dr. J. Cohn Minerva Center for Biomembrane Research (N.D.), and the European Commission (N.D. and I.G.). N.D. is the incumbent of the Philip M. Klutznick professional chair of Development Biology.

² Correspondence: Irit Granot, Department of Biological Regulation, The Weizmann Institute of Science, 76100 Rehovot, Israel. FAX: 972 8 9344116; granotie{at}inter.net.il

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