Limits...
Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture.

Park JW, Shin YK, Choen YP - (2014)

Bottom Line: It showed peak at 2-cell stage and decreased continuously until morula stage.Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture.It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Biology and Physiology, School of Biosciences and Chemistry, Institute for Basic Sciences, Sungshin University, Seoul 147-742, Korea.

ABSTRACT
Adaptive development of early stage embryo is well established and recently it is explored that the mammalian 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 compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second midpreimplantation gene 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-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

No MeSH data available.


Relative Aqp5 mRNA levels in preimplantation stage embryos which were cultured in vitro. Realtime RT-PCR was used to determine the relative mRNA levels of Aqp5 in 4-cell (4-C), 8-cell (8-C), morula (Mor), and blastocyst (Bla) which were cultured in vitro from 2-cell stage. Ten embryos were used for real-time RT-PCR per embryonic stage and repeated 3 times in all stages. * Significant difference among stages (p<0.05, ANOVA).
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC4282210&req=5

Figure 2: Relative Aqp5 mRNA levels in preimplantation stage embryos which were cultured in vitro. Realtime RT-PCR was used to determine the relative mRNA levels of Aqp5 in 4-cell (4-C), 8-cell (8-C), morula (Mor), and blastocyst (Bla) which were cultured in vitro from 2-cell stage. Ten embryos were used for real-time RT-PCR per embryonic stage and repeated 3 times in all stages. * Significant difference among stages (p<0.05, ANOVA).

Mentions: Under the BWW, 37°C, 5% CO2 in > 95% humidified air culture condition, the Aqp5 transcript was detected in all the stages examined. Aqp5 mRNA expression patterns were similar to in vivo, but relative fold change was higher than in vivo (Fig. 2). It means that the expression patterns were not changed both in Aqp5 expression but the levels of expression or degradation were modified by in vitro culture.


Adaptive Transition of Aquaporin 5 Expression and Localization during Preimplantation Embryo Development by In Vitro Culture.

Park JW, Shin YK, Choen YP - (2014)

Relative Aqp5 mRNA levels in preimplantation stage embryos which were cultured in vitro. Realtime RT-PCR was used to determine the relative mRNA levels of Aqp5 in 4-cell (4-C), 8-cell (8-C), morula (Mor), and blastocyst (Bla) which were cultured in vitro from 2-cell stage. Ten embryos were used for real-time RT-PCR per embryonic stage and repeated 3 times in all stages. * Significant difference among stages (p<0.05, ANOVA).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4282210&req=5

Figure 2: Relative Aqp5 mRNA levels in preimplantation stage embryos which were cultured in vitro. Realtime RT-PCR was used to determine the relative mRNA levels of Aqp5 in 4-cell (4-C), 8-cell (8-C), morula (Mor), and blastocyst (Bla) which were cultured in vitro from 2-cell stage. Ten embryos were used for real-time RT-PCR per embryonic stage and repeated 3 times in all stages. * Significant difference among stages (p<0.05, ANOVA).
Mentions: Under the BWW, 37°C, 5% CO2 in > 95% humidified air culture condition, the Aqp5 transcript was detected in all the stages examined. Aqp5 mRNA expression patterns were similar to in vivo, but relative fold change was higher than in vivo (Fig. 2). It means that the expression patterns were not changed both in Aqp5 expression but the levels of expression or degradation were modified by in vitro culture.

Bottom Line: It showed peak at 2-cell stage and decreased continuously until morula stage.Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture.It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

View Article: PubMed Central - PubMed

Affiliation: Division of Developmental Biology and Physiology, School of Biosciences and Chemistry, Institute for Basic Sciences, Sungshin University, Seoul 147-742, Korea.

ABSTRACT
Adaptive development of early stage embryo is well established and recently it is explored that the mammalian 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 compare expression patterns between in vivo and in vitro, we conducted quantitative RT-PCR and analyzed localization of the AQP5 by whole mount immunofluorescence. At in vivo condition, Aqp5 expressed in oocyte and in all the stages of preimplantation embryo. It showed peak at 2-cell stage and decreased continuously until morula stage. At in vitro condition, Aqp5 expression pattern was similar with in vivo embryos. It expressed both at embryonic genome activation phase and second midpreimplantation gene 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-cell stage from cytoplasm to nucleus by in vitro development. Those results explore the modification of Aqp5 expression levels and location of its final products by in vitro culture. It suggests that expression of Aqp5 and the roles of AQP5 in homeostasis can be modulated by in vitro culture, and that early stage embryos can develop successfully by themselves adapting to their condition through modulation of the specific gene expression and localization.

No MeSH data available.