Nuclear Modeling and Developmental Potential of Bovine Somatic Nuclear Transfer Embryos Cloned by Two Different Activation Methods

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 1호
ㆍ저자명 : Byeong-Gyun Jeon, Gyu-Jin Rho

목차

ABSTRACT INTRODUCTION MATERIALS AND METHODS Media and Chemicals Preparation of Donor Cells Preparation of Recipient Cytoplasms Nuclear Transfer and Oocyte Activation In Vitro Culture of Embryos Immunofluorescent Staining of Embryos Absolutely Quantitative Analysis of Transcripts by RealTime RT-PCR Relative-quantitative Telomerase Repeat AmplificationProtocol (RQ-TRAP) Statistical Analysis RESULTS Developmental Potential of SCNT Embryos Relative Telomerase Activity (RTA) of SCNT Embryos Transcript Levels of X-linked Genes in SCNT Embryos DISCUSSION ACKNOWLEDGEMENTS REFERENCES

영어 초록

The present study investigated the nuclear remodeling, development potential with telomerase activity and transcription level of X-linked genes (ANT3, HPRT, MeCP2, RPS4X, XIAP, XIST and ZFX) in the bovine somatic cell nuclear transfer (SCNT) embryos using two different fusion and activation methods. Female adult fibroblasts were injected into perivitelline space of in vitro matured oocytes. The oocyte-nucleus complexes were fused and followed by immediately either activated (Group 1), or activated at 1 h post-fusion (hpf) (Group 2), respectively. The incidence of normal premature chromosome condensation (PCC) at 1 hpf was slightly increased in the Group 2, compared to those of Group 1, but there was no significant (p<0.05) difference. The incidence of normal pronucleus (PN) and chromosome spread at 5 and 18 hpf were significantly (p<0.05) higher in the Group 2 than those of Group 1. The cleavage rate to 2-cell stage, developmental rate to blastocyst stage, and the mean number of total and ICM cell numbers were significantly (p<0.05) higher in the Group 2, compared to those of Group 1. Level of telomerase activity was significantly (p<0.05) higher in the SCNT blastocysts of Group 2, compared to those of Group 1. Transcript levels of HPRT, MeCP2 and XIST were not significantly (p<0.05) different between blastocysts of Group 1 and 2. However, transcript level of ANT3, RPS4X, XIAP and ZFX were significantly (p<0.05) up-regulated in the SCNT blastocysts of Group 2, compared to those of Group 1. Taken together, it is concluded that oocyte activation at 1 hpf induces the enhanced developmental potential by efficient nuclear remodeling and subsequent facilitation of the nuclear reprogramming of bovine SCNT embryos.

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