Effect of cell-penetrating peptide-conjugated estrogen-related receptor beta on the development of mouse embryos cultured in vitro / 대한생식의학회지
Clinical and Experimental Reproductive Medicine
; : 1-8, 2014.
Article
em En
| WPRIM
| ID: wpr-50505
Biblioteca responsável:
WPRO
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.
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Texto completo:
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Índice:
WPRIM
Assunto principal:
Oócitos
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Blastocisto
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RNA Mensageiro
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Imuno-Histoquímica
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Contagem de Células
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Western Blotting
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Desenvolvimento Embrionário
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Estruturas Embrionárias
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Estrogênios
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Receptores Nucleares Órfãos
Limite:
Animals
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Pregnancy
Idioma:
En
Revista:
Clinical and Experimental Reproductive Medicine
Ano de publicação:
2014
Tipo de documento:
Article