The effect of cilostamide on gap junction communication dynamics, chromatin remodeling, and competence acquisition in pig oocytes following parthenogenetic activation and nuclear transfer

Biol Reprod. 2013 Sep 27;89(3):68. doi: 10.1095/biolreprod.113.110577. Print 2013 Sep.


In the pig, the efficiency of in vitro embryo production and somatic cell nuclear transfer (SCNT) procedures remains limited. It has been suggested that prematuration treatments (pre-IVM) based on the prolongation of a patent, bidirectional crosstalk between the oocyte and the cumulus cells through gap junction mediate communication (GJC), with the maintenance of a proper level of cAMP, could improve the developmental capability of oocytes. The aim of this study was to assess: 1) dose-dependent effects of cilostamide on nuclear maturation kinetics, 2) the relationship between treatments on GJC functionality and large-scale chromatin configuration changes, and 3) the impact of treatments on developmental competence acquisition after parthenogenetic activation (PA) and SCNT. Accordingly, cumulus-oocyte complexes were collected from 3- to 6-mm antral follicles and cultured for 24 h in defined culture medium with or without 1 μM cilostamide. GJC functionality was assessed by Lucifer yellow microinjection, while chromatin configuration was evaluated by fluorescence microscopy after nuclear staining. Cilostamide administration sustained functional coupling for up to 24 h of culture and delayed meiotic resumption, as only 25.6% of cilostamide-treated oocytes reached the pro-metaphase I stage compared to the control (69.7%; P < 0.05). Moreover, progressive chromatin condensation was delayed before meiotic resumption based upon G2/M biomarker phosphoprotein epitope acquisition using immunolocalization. Importantly, cilostamide treatment under these conditions improved oocyte developmental competence, as reflected in higher blastocyst quality after both parthenogenetic activation and SCNT.

Keywords: PDE3; SCNT; chromatin; cumulus cells; cyclic adenosine monophosphate (cAMP); embryonic development; gamete biology; gap junctions; meiosis; oocyte; parthenogenetic activation; phosphodiesterases; porcine/pig.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Communication / drug effects*
  • Cells, Cultured
  • Chromatin Assembly and Disassembly / drug effects*
  • Cumulus Cells / drug effects
  • Cumulus Cells / physiology
  • Dose-Response Relationship, Drug
  • Female
  • Gap Junctions / drug effects*
  • Meiosis / drug effects
  • Nuclear Transfer Techniques / veterinary*
  • Oocytes / drug effects*
  • Oocytes / physiology
  • Oogenesis / drug effects
  • Parthenogenesis / drug effects*
  • Parthenogenesis / physiology
  • Quinolones / pharmacology*
  • Swine / physiology*


  • Quinolones
  • cilostamide