Factors Affecting Mouse Somatic Cell Nuclear Reprogramming by Rabbit Ooplasms

Cell Reprogram. 2017 Dec;19(6):344-353. doi: 10.1089/cell.2017.0021. Epub 2017 Nov 14.

Abstract

Successful development of interspecies somatic cell nuclear transfer (iSCNT) embryos depends on compatibilities between ooplasmic and nuclear components. However, the mechanisms by which the compatibilities are regulated are still unknown. In this study, using mouse Oct4-green fluorescent protein (GFP) cells as donors and rabbit oocytes as recipients, we show that Oct4 and other pluripotency related genes were reactivated in some of mouse-rabbit iSCNT embryos, which could also activate Oct4 promoter-driven GFP reporter gene expression. Series nuclear transfer improved the efficiency of Oct4 reactivation. DNA demethylation of Oct4 promoter was detected in GFP positive iSCNT blastocysts, whereas GFP negative iSCNT embryos showed a low efficiency. Our results demonstrate that Oct4-GFP can well label the embryos with epigenetic remodeling and reactivation of pluripotent gene expression. Abundant rabbit mitochondria specific DNAs were identified in reconstructed mouse-rabbit embryos throughout preimplantation stages. Our data demonstrate that epigenetic remodeling and the complete mitochondrial match are not necessary for successful iSCNT embryo development before implantation.

Keywords: DNA demethylation; Oct4; embryonic stem cells; interspecies nuclear transfer; reprogramming.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / physiology
  • Cellular Reprogramming*
  • Cloning, Organism
  • Cytoplasm
  • DNA Methylation
  • DNA, Mitochondrial / genetics
  • DNA, Mitochondrial / metabolism
  • Embryo, Mammalian / cytology
  • Embryo, Mammalian / physiology*
  • Embryonic Development
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / physiology*
  • Epigenesis, Genetic*
  • Female
  • Green Fluorescent Proteins / metabolism
  • Hybrid Cells
  • Mice
  • Nuclear Transfer Techniques / veterinary*
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Oocytes / cytology
  • Oocytes / physiology*
  • Rabbits

Substances

  • DNA, Mitochondrial
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • Green Fluorescent Proteins