Post-activation treatment with demecolcine improves development of somatic cell nuclear transfer embryos in pigs by modifying the remodeling of donor nuclei

Mol Reprod Dev. 2009 Jul;76(7):611-9. doi: 10.1002/mrd.20989.

Abstract

The objective of this study was to examine the effect of cytochalasin B (CB) and/or demecolcine (Dc) on the remodeling of donor nuclei, nuclear ploidy, and development of somatic cell nuclear transfer (SCNT) and parthenogenetic (PA) pig embryos. SCNT and PA oocytes were either untreated (control), or treated with CB, Dc, or both CB and Dc after electric activation, and then cultured or transferred to surrogates. In SCNT, blastocyst formation was higher after treatment with CB and/or Dc (26-28%) than in the controls (16%). The number of oocytes that formed a single pronucleus (PN) was higher after treatment with Dc (86%) and CB + Dc (86%) than under control conditions (44%) or after treatment with CB (63%). In PA, blastocyst formation was higher after CB treatment (47%) than under control conditions (28%), while the formation of a single PN was higher after treatment with Dc (88%) and CB + Dc (84%) compared to controls (34%). The rate of formation of diploid embryos was higher after treatment with Dc and CB + Dc than under control conditions. Dc treatment resulted in a farrowing rate of 50% with 1.1% production efficiency, while controls showed a farrowing rate of 37.5% and a production efficiency of 0.7%. The results of our study demonstrate that post-activation treatment with Dc improves preimplantation development and supports normal in vivo development of SCNT pig embryos, probably because Dc induces formation of a single PN and this leads to normal nuclear ploidy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Nucleus / drug effects*
  • Cells, Cultured
  • Chromatin / drug effects
  • Cytochalasin B / pharmacology
  • Data Interpretation, Statistical
  • Demecolcine / pharmacology*
  • Embryo, Mammalian / drug effects
  • Nuclear Transfer Techniques*
  • Oocytes / drug effects*
  • Oocytes / metabolism
  • Parthenogenesis
  • Ploidies
  • Swine
  • Tubulin / metabolism
  • Tubulin Modulators / pharmacology*

Substances

  • Chromatin
  • Tubulin
  • Tubulin Modulators
  • Cytochalasin B
  • Demecolcine