Purification Of Pmsg And Its Effect On Ovary And Uterus Of Immature Mice And Pre-puberal Blue Foxes

Pregnant mare serum gonadotropin(PMSG) is a glycoprotein hormone,which is produced by trophoblast cells of endometrial cups in pregnant mares and plays an important role in follicle development,ovulation and corpus luteum formation.Using the methods of precipitation with metaphosphoric acid(HPO₃) and fractionated precipitation with alcohol,the crude PMSG was extracted from the pregnant mare serum.Its bioactivity was tested through examining effect of the extracted PMSG on female reproductive organ development of mice and rats.Beside that,the effect of PMSG combined with E2,PGF2a and hCG on induction of estrus,ovarian and follicular development of the month 6 to 7 female blue fox was also tested. Main results and conclusion were showed as follows:(1) Crude activity proteins were exstracted(total wet weight,116.15g) from 9000ml pregnant mare serum.The specific activity of the crude PMSG was between 104.57 and 222.04 IU/mg and the mean specific activity was 153 IU/mg.It indicated that the method was suit for extracting PMSG from pregnant mare serum.(2) Crude proteins were purified using anion-exchange chromatography in a batch-wise mode and cation-exchange column chromatography.The specific activity of the purified PMSG was 173.08 IU/mg and 926.41 IU/mg.It suggested that the purified PMSG could be obtained by ion-exchange chromatography.(3) Three doses of PMSG(2IU,4IU and 8IU) were injected into female mice of postnatal day 21 respectively,and the result showed that 4 IU of PMSG was appropriate to promote mice ovarian development.It indicated that the development of ovary was significantly promoted by treating with a suitable concentration of PMSG.(4) After PMSG injecting to the mice of different ages(21d,15d,10d,and 5d),PMSG promoted not only development of antral follicles in the treatment group of 21d and 15d,but also that of secondary follicles of postnatal day 21 mice.PMSG could promote the postnatal day 15 to day 21 mice uterine growth.On the contrary,PMSG could not promote the postnatal day 5 and day 10 mice uterine growth.(5) Month 6 to 7 female foxes were inducted estrus by E₂+PGF₂a+PMSG and E₂+PGF₂a+ PMSG,and the rate of estrus were 20%and 30%respectively.However,effects of the hormone were heterogenous on different fox.Moreover,estrus of female foxes could be induced by those treatments.(6) The weight of ovary and the amount of antral follicles were increased by injecting E₂+ PGF₂a+PMSG and E₂+PGF₂a+PMSG+hCG.On the other hand,the development of uterus was also significantly promoted But the amount of atresic and abnormal follicles were also increased.It indicated the development of reproductive organ of pre-puberal blue foxes could be promentd by this treatment method.These results could further explain how the PMSG affect the development of the reproductive organs of the fur animal during the estrus and non-estrus periods,and it could help to establish PMSG application procedure,develop its purification skills and produce PMSG antiserum in the future.