Evaluation Of The Effect Of Premature Ovarian Failure(POF)Transgenic Mouse Model

As the rhythm of the modern society has accelerated and the pressure has increased,reproductive diseases have become a serious disease that afflicts humans.Among them,for women,premature ovarian failure(POF)can cause women suffering from infertility.And the incidence of premature ovarian failure is increasing with the age of women.Among young women under the age of 20,the incidence of premature ovarian failure is 1/10,000.When women reach the age of 30,the incidence of premature ovarian failure increases.To 1/1000.When women reach the age of 40,the incidence of premature ovarian failure is 1%.Once infertility is created,women’s reproductive ability cannot be restored.And so far there is no particularly effective treatment.With the discovery of germline stem cells and the maturation of culture techniques,germline stem cell transplantation has become an attempted therapy.Because the mouse has a short reproductive cycle and is easy to operate,mice were selected as model animals to obtain Kunming-transgenic p Sohlh2-Tet3G-TRE 3G-Flag-DTA transgenic mice by prokaryotic injection.The expression of diphtheria toxin A(DTA)was induced by doxycycline(Dox)in Tet-on expression system.Under the action of promoter Sohlh2,the germ cells are specifically removed.First of all,the transgenic mice were expanded,then test crossing with wild type Kunming mice,detect the positive rate in offspring;screening homozygous transgenic mice.The female mice were drug-induced and it was found that DOX feeding two weeks before the pregnancy until the offspring mice grew up,can have the best germ cells removal effect.Parallel control experiments were wild-type Kunming mice.After 8 weeks of induction,paraffin sections were prepared from the ovary of female mice,and morphological observation was performed under a 200 X microscope to find that the primordial follicles were clearly eliminated.In order to determine this phenomenon,follicle counting experiments were performed on the ovary of female-induced transgenic mice and wild female mice of the control group.The number of follicles in each stage was statistically analyzed,and it was concluded that the difference in primordial follicles in the ovaries of transgenic female mice and wild female mice in the control group was extremely significant.To further detect the expression of the target protein in the mouse ovary,we used frozen sections to perform in situ hybridization experiments of DTA and rt TA.The results of in situ hybridization experiments showed that two genes,DTA and rt TA,were mainly expressed in the ovarian cortex.The results obtained with the follicle counting experiment were in agreement.In order to detect the growth of transgenic mice,we took 15 6-week-old female transgenic mice and wild-type mice for weight measurement.The results showed that there was no significant difference between the body weight of the transgenic female mice and the wild-type female mice,so the gene transfer did not interfere with the growth of the mice.The estrous cycle of transgenic mice and wild-type mice was then tested to determine the duration of each phase of the estrus cycle.The experimental results showed that there was no significant difference between the estrus cycle of transgenic female mice and the estrous cycle of wild-type female mice,but the overshoot of wild-type mice during each phase of the estrus cycle was more stable.In conclusion: the present experiment demonstrated that the ovary in transgenic female mice was significantly emptied by detecting ovarian morphology,follicle count,estrus cycle and in situ hybridization experiments in transgenic female and wild female mice,laid the foundation for the subsequent experimental study.