Effects of spindle removal on MPF and MAP kinase activities in porcine matured oocytes

Seitaro Goto, Kunihiko Naito, Satoshi Ohashi, Koji Sugiura, Haruna Naruoka, Naoki Iwamori, Hideaki Tojo

研究成果: ジャーナルへの寄稿記事

17 引用 (Scopus)

抄録

Intracellular localization of maturation/M-phase promoting factor (MPF) and mitogen activated protein (MAP) kinase in mature oocytes has been examined by immunocytochemical methods and the authors of these studies have reported that they are localized on spindles during M-phase. Although these reports showed the relative localization of MPF and MAPK on spindles, it has never been shown whether these kinases are present in the cytoplasm and, if they are present, how many parts of the kinases are localized on the metaphase spindle. In the present study, we made quantitative analyses of MPF and MAP kinase localized on oocyte spindles by kinase assays and immunoblotting after removal of the spindles from porcine mature oocytes. First, we certified their intracellular distribution by immunocytochemical methods and observed sharp signals of cyclin B1 on spindle poles and MAP kinase signals on the microtubule of metaphase spindles. In contrast to these results by immunostaining, the amounts of MPF and MAP kinase localized on spindles examined by immunoblotting and kinase assays were undetectable and less than 20%, respectively. These results indicate that the immunocytochemical technique is a powerful method for showing relative localization, but it is not suitable for quantitative analysis, and that the removal of metaphase spindles from mature oocytes does not have a severe negative impact on the subsequent MPF and MAP kinase activity and on the cell cycle progression in early embryo development.

元の言語英語
ページ(範囲)388-393
ページ数6
ジャーナルMolecular Reproduction and Development
63
発行部数3
DOI
出版物ステータス出版済み - 11 1 2002
外部発表Yes

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Maturation-Promoting Factor
Mitogen-Activated Protein Kinases
Oocytes
Swine
Phosphotransferases
Metaphase
Immunoblotting
Spindle Poles
Cyclin B1
Microtubules
Cell Division
Embryonic Development
Cell Cycle
Cytoplasm

All Science Journal Classification (ASJC) codes

  • Genetics
  • Developmental Biology
  • Cell Biology

これを引用

Effects of spindle removal on MPF and MAP kinase activities in porcine matured oocytes. / Goto, Seitaro; Naito, Kunihiko; Ohashi, Satoshi; Sugiura, Koji; Naruoka, Haruna; Iwamori, Naoki; Tojo, Hideaki.

:: Molecular Reproduction and Development, 巻 63, 番号 3, 01.11.2002, p. 388-393.

研究成果: ジャーナルへの寄稿記事

Goto, Seitaro ; Naito, Kunihiko ; Ohashi, Satoshi ; Sugiura, Koji ; Naruoka, Haruna ; Iwamori, Naoki ; Tojo, Hideaki. / Effects of spindle removal on MPF and MAP kinase activities in porcine matured oocytes. :: Molecular Reproduction and Development. 2002 ; 巻 63, 番号 3. pp. 388-393.
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abstract = "Intracellular localization of maturation/M-phase promoting factor (MPF) and mitogen activated protein (MAP) kinase in mature oocytes has been examined by immunocytochemical methods and the authors of these studies have reported that they are localized on spindles during M-phase. Although these reports showed the relative localization of MPF and MAPK on spindles, it has never been shown whether these kinases are present in the cytoplasm and, if they are present, how many parts of the kinases are localized on the metaphase spindle. In the present study, we made quantitative analyses of MPF and MAP kinase localized on oocyte spindles by kinase assays and immunoblotting after removal of the spindles from porcine mature oocytes. First, we certified their intracellular distribution by immunocytochemical methods and observed sharp signals of cyclin B1 on spindle poles and MAP kinase signals on the microtubule of metaphase spindles. In contrast to these results by immunostaining, the amounts of MPF and MAP kinase localized on spindles examined by immunoblotting and kinase assays were undetectable and less than 20{\%}, respectively. These results indicate that the immunocytochemical technique is a powerful method for showing relative localization, but it is not suitable for quantitative analysis, and that the removal of metaphase spindles from mature oocytes does not have a severe negative impact on the subsequent MPF and MAP kinase activity and on the cell cycle progression in early embryo development.",
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