Effect Of AY 9944-A-7 On Porcine Oocytes Maturation In Vitro And The Expression Of C-mos, MEK And ERK1/2

In the pathway of cholesterol metabolism, follicle fluid-meiosis activating sterol (FF-MAS) can be transformed to testis-meiosis activating sterol (T-MAS) by the action of ?14-reductase. AY 9944-A-7 can induce accumulation of FF-MAS in sexual glands by inhibiting the activity of ?14-reductase, thus blocking the transformation of FF-MAS to T-MAS. This study mainly explored the function of AY 9944-A-7 in the maturation of porcine oocytes. The release of first polar body was considered as the maturation of porcine oocytes. Besides, influence of AY 9944-A-7 to the expression of genes in Mos/MEK/ERK1/2 pathways was also investigated. Details are showed below:Cumulus embrace oocytes (CEOs) were treated with AY 9944-A-7 in different concentrations (0,20, 30,40?mol/L) to study the function of AY 9944-A-7 in in vitro maturation of porcine CEOs. Moreover, the influence of AY 9944-A-7 with different concentrations on the expression of key genes in Mos/MEK/ERK1/2 pathways was detected using quantitative real time PCR in matured oocytes. Results showed that,30?mol/L AY 9944-A-7 had best effort on the maturation of pocine CEOs in vitro. C-mos gene had the highest abundance in porcine CEOs cultured with 20?mol/L AY 9944-A-7. Besides, MEK, ERK1 and ERK2 gene had the highest abundance in porcine CEOs cultured with 30?mol/L AY 9944-A-7.Porcine CEOs were cultured in medium with 30?mol/L AY 9944-A-7 for different time (12h,24h, 36h,44h). Harvested oocytes were investigated to find out the influence of different cultured time on the expression of key genes in Mos/MEK/ERK1/2 pathway. Expression of c-mos gene in cells cultured with 30 ?mol/L AY 9944-A-7 for 12h showed at most. MEK had highest abundance after cultured for 12 h and/or 44h, which showed no significant difference (P>0.05). ERK1 and ERK2 all showed highest abundance after cultured for 44 h.Above knowable, the best concentration of AY 9944-A-7 for inducing the porcine CEOs to recover meiosis is 30?mol/L. The concentration of AY 9944-A-7 in medium and/or culturing time can influence the expression of key genes in Mos/MEK/ERK1/2 pathway.