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Factors Affecting Mouse Somatic Cell Nuclear Reprogramming by Rabbit Ooplasms.

Xia Huang1, Lili Song1, Zhiyan Zhan1

  • 11 Key Laboratory of Pediatric Hematology and Oncology Ministry of Health, Pediatric Translational Medicine Institute, Shanghai Children's Medical Center, School of Medicine, Shanghai Jiao Tong University , Shanghai, China .

Cellular Reprogramming
|November 15, 2017
PubMed
Summary
This summary is machine-generated.

Interspecies somatic cell nuclear transfer (iSCNT) requires component compatibility. Mouse-rabbit iSCNT embryos showed gene reactivation and epigenetic remodeling, indicating successful early development without mitochondrial matching.

Keywords:
DNA demethylationOct4embryonic stem cellsinterspecies nuclear transferreprogramming

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Area of Science:

  • Reproductive Biology
  • Developmental Biology
  • Genetics

Background:

  • Successful interspecies somatic cell nuclear transfer (iSCNT) development hinges on ooplasmic-nuclear component compatibility.
  • Mechanisms regulating these compatibilities in iSCNT remain largely unknown.
  • Understanding these factors is crucial for advancing reproductive technologies.

Purpose of the Study:

  • To investigate the reactivation of pluripotency genes, specifically Oct4, in mouse-rabbit iSCNT embryos.
  • To determine the role of epigenetic remodeling, including DNA demethylation, in iSCNT success.
  • To assess the necessity of mitochondrial DNA compatibility for preimplantation iSCNT embryo development.

Main Methods:

  • Utilized mouse Oct4-green fluorescent protein (GFP) cells as nuclear donors and rabbit oocytes as recipients for iSCNT.
  • Employed serial nuclear transfer to enhance the efficiency of Oct4 reactivation.
  • Analyzed Oct4 promoter activity via GFP reporter gene expression and assessed DNA demethylation in resulting blastocysts.
  • Detected and quantified rabbit mitochondria-specific DNA in reconstructed embryos.

Main Results:

  • Oct4 and other pluripotency-related genes were reactivated in some mouse-rabbit iSCNT embryos, evidenced by GFP expression.
  • Serial nuclear transfer improved the efficiency of Oct4 reactivation.
  • GFP-positive iSCNT blastocysts exhibited Oct4 promoter DNA demethylation, correlating with higher developmental efficiency.
  • Abundant rabbit mitochondria DNA was present throughout preimplantation stages in reconstructed embryos.

Conclusions:

  • Oct4-GFP serves as a reliable marker for epigenetic remodeling and pluripotent gene expression reactivation in iSCNT embryos.
  • Complete mitochondrial DNA matching is not essential for successful preimplantation development in this mouse-rabbit iSCNT model.
  • Epigenetic reprogramming plays a significant role in enabling iSCNT embryo development across species.