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KMID : 1033620140410010001
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Clinical and Experimental Reproductive Medicine
2014 Volume.41 No. 1 p.1 ~ p.8
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Effect of cell-penetrating peptide-conjugated estrogen-related receptor β on the development of mouse embryos cultured in vitro
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Yang Ning-Jie
Seol Dong-Won
Jo Jung-Hyun
Jang Hyun-Mee
Yoon Sook-Young
Lee Dong-Ryul
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Abstract
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Objective: Estrogen related receptor β (Esrrb) is a member of the orphan nuclear receptors and may regulate the expression of pluripotency-related genes, such as Oct4 and Nanog. Therefore, in the present study, we have developed a method for delivering exogenous ESRRB recombinant protein into embryos by using cell-penetrating peptide (CPP) conjugation and have analyzed their effect on embryonic development.
Methods: Mouse oocytes and embryos were obtained from superovulated mice. The expression of Oct4 mRNA and the cell number of inner cell mass (ICM) in the in vitro-derived and in vivo-derived blastocysts were first analyzed by real time-reverse transcription-polymerase chain reaction and differential staining. Then 8-cell embryos were cultured in KSOM media with or without 2 μg/mL CPP-ESRRB protein for 24 to 48 hours, followed by checking their integration into embryos during in vitro culture by Western blot and immunocytochemistry.
Results: Expression of Oct4 and the cell number of ICM were lower in the in vitro-derived blastocysts than in the in vivo-derived ones (p<0.05). In the blastocysts derived from the CPP-ESRRB-treated group, expression of Oct4 was greater than in the non-treated groups (p<0.05). Although no difference in embryonic development was observed between the treated and non-treated groups, the cell number of ICM was greater in the CPP-ESRRB-treated group.
Conclusion: Treatment of CPP-ESRRB during cultivation could increase embryos’ expression of Oct4 and the formation rate of the ICM in the blastocyst. Additionally, an exogenous delivery system of CPP-conjugated protein would be a useful tool for improving embryo culture systems.
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KEYWORD
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Estrogen-related receptor β, Cell-penetrating peptide, Oct4, Mouse embryo, In vitro culture
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