Effect Of Trichostatin A On Ovine Oocyte In Vitro

In this study, we investigate the effect of Trichostation A on ovine oocyte in vitro maturation,in vitro fertilization, the embryo anaphase development and the effect of Trichostation A on thepositive oocytes and negative oocytes anaphase development when screened for Brilliant cresylblue, which provide a theoretical basis for further research in the mechanisms of epigeneticregulation and embryonic development. The main findings were as follows:1. Effect of TSA on the development of ovine oocytes and parthenogenetic embryoEmbryo culture medium were added0,50,100,200,300,500nM TSA, we found thatembryo culture was added300nM could significantly improve ovine parthenogenetic embryoblastocyst rate and blastocyst cell number (P<0.05); when300nM TSA treated for0h,5h,15h,we found that treatment with10h could significantly increased blastocyst rate (P<0.05); themature medium would reduce the oocytes maturation and the development of parthenogeneticembryos compared with added TSA when cultured in vitro. Thus, adding300nM TSA to the invitro cultured medium and treated for10h could improve the development of ovineparthenogenetic embryos.2. Effect of TSA on ovine oocytes in vitro maturation and in vitro fertilizationThe different concentrations of TSA (0,50,100,200nM) in vitro ovine oocyte maturationmedium have inhibiting effect on the extension of ovine oocyte. When the concentration was200nM, the maturation and development of oocytes were significantly inhibited. When addeddifferent concentrations of TSA (0,100,200,300nM) in IVF, we found that200nM TSA couldsignificantly improve blastocyst rate and the total number of ovine blastocyst cells of oocyte IVF(P<0.05);200nM TSA treated for0h,10h and24h, Treated with10h could significantlyimprove the blastocyst rate (P<0.05). Thus, the mature medium, added with TSA, has nosignificant effect on the oocytes maturation and the development of embryos. the TSA wouldreduce the oocytes maturation and the development of embryos when concentration increased to200nM. While in vitro fertilization, the200nM TSA treated for10h would significantly improvethe development of fertilization embryo.3. Effect of TSA on the selected by BCB oocytes in development of vitro embryoThe ovine oocytes stained90min in26uM BCB, the oocyte stained cytoplasmic bluemarked with BCB+, cytoplasmic stained no coloration marked with BCB-. The BCB+and BCB-oocytes have parthenogenetic activation and IVF respectively. The results showed that: BCB+blastocyst rate was significantly higher than that in control group and BCB-group in oocytesparthenogenetic activation (P<0.05), BCB-oocyte cleavage rate and blastocyst cell number wassignificantly lower than that in BCB+group. The BCB+oocytes blastocyst rate and blastocystcells were significantly higher than those in control group and BCB-after IVF (P<0.05); the BCB-group blastocyst rate (16.89%vs37.55%) and the total number of blastocyst cell (79.45vs100.29)was significantly lower than the control group (P<0.05). Putting the parthenogenetic embryos andfertilized embryos of BCB-oocytes in300nM TSA embryo culture medium treated with10hwould significantly increased the BCB-oocytes parthenogenetic embryo blastocyst rate (23.33%vs35.65%) and fertilization embryo blastocyst rate (19.36%vs29.46%)(P<0.05); Placing theBCB-oocytes parthenogenetic embryos and fertilization embryos in300nM TSA embryo culturemedium treated with10h does not improve the BCB+oocytes in vitro viability (P>0.05).