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a Department of Animal Science, University of Missouri-Columbia, Columbia, Missouri 65211
Actin is one of the major proteins in mammalian oocytes. Most developmental events are dependent on the normal distribution of filamentous (F-) actin. Polymerization of nonfilamentous (G-) actin into F-actin is important for both meiosis and mitosis. This study examined G- and F-actin distribution in pig oocytes and embryos by immunocytochemical staining and confocal microscopy. Actin protein was quantified by electrophoresis and immunoblotting. G-Actin was distributed in the whole cytoplasm of oocytes and embryos irrespective of their stages. F-Actin was distributed at the cortex of oocytes and embryos at all stages, at the joint of blastomeres in the embryos, in the cytoplasm around the germinal vesicle (GV), and in the perinuclear area of 2- to 4-cell-stage embryos. No differences in the amount of actin protein were found among oocytes and embryos. Oocytes cultured in medium with cytochalasin D (CD), an inhibitor of microfilament polymerization, underwent GV breakdown and reached metaphase I but did not proceed to metaphase II. Two- to 4-cell-stage embryos cultured in medium with CD did not develop to blastocysts. When GV-stage oocytes or 2- to 4-cell-stage embryos treated with CD for 6 h were re-cultured in media without CD, oocytes or embryos re-assembled actin filaments and underwent a meiotic maturation or blastocyst formation similar to that of controls. These results indicate that it is the polymerization of G-actin into F-actin, not actin protein synthesis, that is important for both meiosis and mitosis in pig oocytes and embryos.
1 This research is supported in part by the National Cooperative Program on Non-Human In Vitro Fertilization and Preimplantation Development, funded by the National Institutes of Child Health and Human Development, NIH, through cooperative agreement HD34588. This manuscript is a contribution from the Missouri Agricultural Experiment Station, Journal Series Number 12,896.
2 Correspondence: Billy N. Day, 159 Animal Sciences Research Center, Department of Animal Science, University of Missouri-Columbia, Columbia, MO 65211. FAX: 573 884 7827; dayb{at}missouri.edu
3 Current address: Division of Reproductive Medicine and Infertility, Women & Infants' Hospital of Rhode Island, Brown University School of Medicine, Providence, RI 02905.
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