Study On ET-1 Inducing Of The Mouse Oocyte Maturation In Vitro And It's Mechanism

OBJECTIVEThe meiotic maturation and fertilization of oocytes are central affairs of animal reproduction. ET-1 is one of the important regulation factors in the ovary , and can regulate follicle development, follicle ovulation, corpus luteum formation , luteolysis by autocrine and paracrine, but the mechanism is still not well-known;we studyed the effect of ET-1 on the mouse oocyte maturation in vitro and it's mechanism and hoped it can bring us some new methods for fertility and antifertility.METHODS1.Preparing animal model,harvesting-culturing COC and DOWe used exogenous PMSG to induce Kunming mice superovulate which were wighted about 15 ～ 18g. After 48h , we executed the mouse by decapitation , took out of hibateral ovaries , and lacerate egg by needle under the anatomical microscope . We choosed the COC which is enclosed by 3-5 layers GC. To obtain DO, we used mouth haustorial tube to blow COC . Then we divided oocytes into COC and DO, and observed ET-1 or other factors on oocytes maturation in vitro , GVBD indicates the maturation of oocyte nucleus and the extrusion of PB1 indicates the maturation of oocyte kytoplasm.2. RNA isolation and measuring the expression of c-erbB₂ mRNA and c-myb mRNA in COC by RT-PCR300 COC→adding 0.5 ml denaturing solution, on ice , 5min→add 0.1ml chloroform mixture, shake 15s→4℃,centrifuge at 10000rpm, 20 min→...