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Clin Exp Reprod Med. 2014 Mar;41(1):1-8. doi: 10.5653/cerm.2014.41.1.1. Epub 2014 Mar 14.

Effect of cell-penetrating peptide-conjugated estrogen-related receptor β on the development of mouse embryos cultured in vitro.

Clinical and experimental reproductive medicine

Ning Jie Yang, Dong-Won Seol, Junghyun Jo, Hyun Mee Jang, Sook-Young Yoon, Dong Ryul Lee

Affiliations

  1. Department of Biomedical Science, College of Life Science, CHA University, Seoul, Korea.
  2. Department of Biomedical Science, College of Life Science, CHA University, Seoul, Korea. ; Fertility Center of CHA Gangnam Medical Center, CHA University, Seoul, Korea.

PMID: 24693491 PMCID: PMC3968251 DOI: 10.5653/cerm.2014.41.1.1

Abstract

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.

Keywords: Cell-penetrating peptide; Estrogen-related receptor β; In vitro culture; Mouse embryo; Oct4

References

  1. Nature. 1997 Aug 21;388(6644):778-82 - PubMed
  2. Genes Dev. 1992 Jun;6(6):939-52 - PubMed
  3. Cell. 1988 Dec 23;55(6):1189-93 - PubMed
  4. Hum Reprod. 2008 Dec;23(12):2782-90 - PubMed
  5. Am J Obstet Gynecol. 1991 Dec;165(6 Pt 1):1802-5 - PubMed
  6. Proc Natl Acad Sci U S A. 1990 Jun;87(12):4756-60 - PubMed
  7. J Biol Chem. 2008 Dec 19;283(51):35825-33 - PubMed
  8. Pharm Res. 2007 Nov;24(11):1977-92 - PubMed
  9. Cell. 1998 Oct 30;95(3):379-91 - PubMed
  10. Nature. 1988 Jan 7;331(6151):91-4 - PubMed
  11. Stem Cells. 2012 Aug;30(8):1703-13 - PubMed
  12. Fertil Steril. 1994 Sep;62(3):555-8 - PubMed
  13. Dev Reprod. 2013 Mar;17(1):9-16 - PubMed
  14. Nat Genet. 2000 Apr;24(4):372-6 - PubMed
  15. Mol Reprod Dev. 1992 Mar;31(3):195-9 - PubMed
  16. J Reprod Fertil. 1995 Nov;105(2):331-8 - PubMed
  17. Reprod Fertil Dev. 2010;22(1):75-87 - PubMed
  18. Am J Obstet Gynecol. 1994 Aug;171(2):406-9 - PubMed
  19. Fertil Steril. 1993 Jan;59(1):138-42 - PubMed
  20. J Clin Endocrinol Metab. 2006 Feb;91(2):569-79 - PubMed
  21. J Pharm Sci. 2003 Sep;92(9):1793-804 - PubMed
  22. Mol Hum Reprod. 1998 Nov;4(11):1021-31 - PubMed
  23. Mol Cell Biol. 2008 Oct;28(19):5986-95 - PubMed
  24. Nature. 2006 Aug 3;442(7102):533-8 - PubMed
  25. J Exp Zool. 1991 Apr;258(1):34-47 - PubMed

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