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논문검색

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

초록

영어

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.

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

저자정보

  • Byeong-Gyun Jeon OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Institute of Animal Medicine
  • Gyu-Jin Rho OBS/Theriogenology and Biotechnology, College of Veterinary Medicine, Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Korea

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