Effect of cell-penetrating peptide-conjugated estrogen-related receptor beta on the development of mouse embryos cultured in vitro.
- Author:
Ning Jie YANG
1
;
Dong Won SEOL
;
Junghyun JO
;
Hyun Mee JANG
;
Sook Young YOON
;
Dong Ryul LEE
Author Information
- Publication Type:In Vitro ; Original Article
- Keywords: Estrogen-related receptor beta; Cell-penetrating peptide; Oct4; Mouse embryo; In vitro culture
- MeSH: Animals; Blastocyst; Blotting, Western; Cell Count; Embryonic Development; Embryonic Structures*; Estrogens; Female; Immunohistochemistry; Mice*; Oocytes; Orphan Nuclear Receptors; Pregnancy; RNA, Messenger
- From:Clinical and Experimental Reproductive Medicine 2014;41(1):1-8
- CountryRepublic of Korea
- Language:English
- Abstract: OBJECTIVE: Estrogen related receptor beta (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 microg/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.
