Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture
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Adaptive development of early stage embryo is well established and recently it is explored that themammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin(AQP) 5 gene which is detected during early development were examined by the environmental condition. To compareexpression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantationembryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expressionpattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second midpreimplantationgene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos,and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cellstage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levelsand location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasiscan be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to theircondition through modulation of the specific gene expression and localization
Abstract
Adaptive development of early stage embryo is well established and recently it is explored that themammalian embryos also have adaptive ability to the stressful environment. However, the mechanisms are largely unknown. In this study, to evaluate the possible role of aquaporin in early embryo developmental adaptation, the expression of aquaporin(AQP) 5 gene which is detected during early development were examined by the environmental condition. To compareexpression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantationembryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expressionpattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second midpreimplantationgene activation phase, but the fold changes were modified between in vivo embryos and in vitro embryos. During in vivo development, AQP5 was mainly localized in apical membrane of blastomeres of 4-cell and 8-cell stage embryos,and then it was localized in cytoplasm. However, the main localization area of AQP5 was dramatically shifted after 8-cellstage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levelsand location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasiscan be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to theircondition through modulation of the specific gene expression and localization
참고문헌 (25)
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