Oxford University Press, Biology of Reproduction, 3(89), 2013
DOI: 10.1095/biolreprod.113.110577
Full text: Download
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), together with the maintenance of a proper level of cAMP, could improve the developmental capability of oocytes. The aim of this study was to assess: (i) dose dependent effects of cilostamide on nuclear maturation kinetics; (ii) the relationship between treatments on GJC functionality and large-scale chromatin configuration changes; (iii) and the impact of treatments on developmental competence acquisition after parthenogenic activation (PA) and SCNT. Accordingly, COC were collected from 3-6 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 up to 24 h of culture and delayed meiotic resumption as only 25.6% of cilostamide-treated oocytes reached ProMI 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.