Effect of phosphodiesterase type 3 inhibitor, cilostamide, on the developmental competence of ovine oocytes isolated by glucose 6-phosphate dehydrogenase activity

Azari, V.; Rahmani, H.; Hajian, M.; Ostadhosseini, S.; Hosseini, S.; Nasr-Esfahani, M.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p20
Academic Journal
Introduction: The developmental competence of oocytes matured in vitro (IVM) is yet far below than in vivo counterparts. Recent studies suggest that the asynchrony between nuclear/cytoplasmic maturation and the initial low/heterogeneous quality of oocytes are the most important factors affecting IVM success. We investigated whether selection of growing oocytes (based on their glucose 6- phosphate dehydrogenase (G6PDH) activity) and chemical induction of nuclear/cytoplasmic synchrony (through transient inhibition of meiosis resumption with cilostamide) can improve developmental competence of sheep oocytes. Materials and Methods: Abattoir-derived oocytes were stained with 26uM BCB for 45 minutes to isolate growing (high G6PDH activity) and fully grown (low G6PDH) oocytes according to their differential capacities in breaking BCB and retaining colorless (BCB-) or blue (BCB+) ooplasm, respectively. BCB-and BCB+ oocytes were then incubated with 1 ng/ml cilostamide for 6h before culture in normal IVM medium. Matured oocytes were then used for embryo development assessment using parthenogenesis activation. Results: Cilostamide delayed meiotic progression in BCB- ovine oocytes. The cleavage, blastocyst and hatching rates in BCB- oocytes that treated by cilostamide were higher than the control group, although these increases were not statistically significant. Conclusion: We concluded that increase of ovine oocyte cAMP concentration during two-step culture partially improves yield and quality of in vitro embryo. This also may suggest that phosphodiesterase type 3-mediated inhibition of cAMP activity is not the only mechanism that controls the process of nuclear maturation in ovine oocyte.


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