The Role And Regulatory Mechanisim Of AL-8810011Oocyte In Vitro Maturation And Development Of Early Embryo In

75-80%of cow’s embryo loss is caused by early embryo death, more than40%of the embryo loss occurs in8-17days of the early pregnancy. Many researches had proved that early pregnancy was bound up with secretion of Prostaglandin F2alpha(PGF2α) in the organism. From in vivo and in vitro studies, our research used PGF2α receptor antagonist AL8810to study effects of buffaloes’ conception rate, luteal function and morphology, development of oocytes and early embryos. Try to investigate the role and regulation mechanism of buffalo oocyte and early embryo development. It may have significance on reducing early embryonic loss and improving the rate of reproduction. The main results are shown as follows:1. Expression of PTGFR mRNA in different developmental stages of buffalo IVF embryosOur results suggest that PTGFR mRNA was expressed in all the different developmental stages of buffalo IVF embryos. It showed a significant rise in4-16cell stage compared with the ova nuda period(P＜0.01). But when it came to the morula stage, the expression of PTGFR mRNA was significantly lower than4-16cell stage (P＜0.01).2. Immunofluorescence decetion of PTGFR protein in Buffalo oocyte and IVF embryoOur results suggest that there were signals for immunofluorescence decetion of PTGFR protein on chromosomes of Buffalo oocyte and IVF embryo.3. Effect of AL-8810on buffalo oocytes in vitro maturation and apoptosisDifferent concentration (100nM,200nM,500nM,800Nm,1600nM) of AL-8810had no significant effect on buffalo oocyte in vitro nuclear maturation (P>0.05). The rate of early apoptosis cells in high-dose treatments (1600nM) was significantly higher than that of the control treatment(63.57%vs.27.02%, P＜0.05). Late apoptosis rates of each treatment were lower than those of control(P＜0.05), while late apoptosis rate was only7.35%of the high dose treatment (1600nM) and which was significantly lower than control(P＜0.05). On the contrary, the rate of morula in high-dose treatments (800nM,1600nM) was significantly lower than that of the low-dose group(100nM)(P＜0.05) in the test of oocyte developmental competence. 4. Effect of AL-8810on buffalo IVF embryosDuring different treatment stages (0-48h\48-96h\96-144h), concentration of AL-8810(100nM\200nM\400nM\800nM\1600nM) had significant effect on the16-cell stage embryos (P=0.003, P=0.004respectively). The treatments in0-48h and48-96h were significantly better than that in96-144h (70.09%,73.43%vs.56.76%) and control, and the optimal treatment combination were0-48h-800nM(88.67%) and48-96h-400nM (85.00%). In addition, the rates of morula in those groups treated with AL-8810before96h, were up regulated, but not significantly (P>0.05), and the the morula rates of48-96h-800nM (50.00%) was higher than other groups (P<0.05). The rates of blastocyst with AL-8810treated before96h were higher than the one after (3.61%,2.78%vs.1.39%)(P>0.05). And AL-8810treatment can inhibit the buffalo IVF embryos PTGFR mRNA expression in a concentration-dependent manner. There were significant differences (P<0.05) between high-dose treatments (400nM,800nM,1600nM) and control group (OnM).5. Effect of AL-8810treatment on morphology and founction of buffalo corpus luteumAL-8810diluent (5000nM) was delivered into uterus opening by the method of artificial insemination. We found this AL-8810treatment had no notable impact on diameter of buffalo corpus luteum(P＞0.05), so as the main hormone in blood(LH\E2\P4)(P＞0.05).6. Effect of AL-8810treatment on Buffalo conception rateIntrauterine injection of AL-8810(5000nM0.25ml), after artificial insemination, had no notable impact on conception rate of Buffalo.In conclusion, PTGFR mRNA expressed both in oocytes and embryos. Moreover, the PGF2a receptor antagonist AL-8810could reduce the expression of PTGFR mRNA, increase the rate of16-cell stage embryos as well. But the effect of AL-8810on buffalo corpus luteum and embryo in voivo are needed a deeply study.