| Objective:To investigate the role of Activin A and Inhibinαin the mice endometrium during blastocyst implantation.Methods:We took mature and nulliparous kunming female mice as subjectives two kunming adult female mice were caged with a normal or a vasoligation male overnight.In the following morning,females were checked for the presence of a vaginal sperm plug.The day vaginal plug was regarded as day 1 of pregnancy or pseudopregnancy.Firstly,pregnanct mice and pseudopregnant mice of day 2-6 were put todeath through cervical vertebrate luxation,and then the uteri were disemboweled and the unpregnant mice uteri were(estrous stage)taken as controls.The uteri were fixed in 4% paraformaldehyde.Hematoxylin-Eosin(HE) staining was used to observe histological structures in the endometrium of pregnant and paeudopregnant mice;Ultrastructure changes in the endometrium of pregnant mice were observed by transmission electron microscope(TEM);Immunhistochemical SP staining was ueded to measure the expression of Activin A and Inhibinα,respectively.Pictures were taken by microscopic imaging systemand then the results of immunohistochemical staining were analysed by computer.To analyze the influence of Activin A and Inhibinαin embryonic implantation,the number of blastocyst implantation was registered on pregnant day 8 after mice being injected Activin A antibody in horn of uterus at 9AM on the pregnant day 4.Result:1.HE staining showed that structural changes of mice endometrium exhibited the developmental tendency on day 2-6 of pregnancy.The amounts uterine glands increased gradually,and the endometrium showed physiological edema.Moreover,the intercellular space of stroma cells became augmented,and blood vessels proliferation.The most obvious changes appeared on day 4-5 of pregnant mice;2.Under TEM,cell organs in luminal epithelium,glandular epithelium and stroma cells in endometrium on day 2-6 of pregnancy were more than those in unpregnant mice(estrous stage).Abundant Mi,RER,SG and so on in luminal epithelium and glandular epithelium and interstitial edema were observed on day 4-5 of pregnancy.Moreover,stroma cells got hypertophy,abound with affluent glycogen and lipid droplet intracytoplasm;3.The results of immunohistochemisty revealed that the expressions of the Activin A were distribute differently in the endometrial luminal,glandual epithelium and stromal cells of unpregnant(estrous stage),pregnant and some pseudopregnant mice.The expression of the Activin A reached the peak on day 4-5 of pregnant mice.The Inhibinαwas expressed in the endometrial luminal,glandual epithelium cells of unpregnant(estrous stage),pregnant and pseudopregnant mice,and the most Inhibinαappeared on day 4-5.The expressions of Activin A and Inhibinαin pseudopregnant mice were differentially from the pregnant mice.4.The mean optical densities(MOD) of Activin A and Inhibinαin pregnant mice on day 4-5 were calculated through image analysis.The data by SPSS software indicated that there was a positive correlation between the expression of Activin A and Inhibinαin the peri-implantation endometrium of pregnant mice.5.To analyze the influence of Activin A in embryonic implantation,the number of blastocyst implantation was registered on pregnant day 98after mice being injected Activin A antibody in horn uterus at 9AM on the pregnant day 4.Conclusion:1.The morphological changes in the endometrium of pregnant mice exhibited a dynamic changing tendency and prominent changes appeared during the day 4-5 of pregnancy.2.The continued expression of Activin A in the endometrial luminal and glandual epithelium and stromal cells of pregnant mice on day 2-6 suggested that Activin A might play an important role in the process of the embryo implantation in mouse.3.The continued expression of Inhibinαin the endometrial luminal and glandual epithelium cells of pregnant mice on day 2-6 suggested that Inhibinαmight play an important role in the process of the embryo implantation in mouse.4.In the endometrium of pseudopregnant mice,the law revealed that the expressions of Activin A and Inhibinαwere regulated not only by matermal normonr,but also by the implanted embryos and other chemical signals.5.The expressions of Activin A and Inhibinαhave a positive correlation in the peri-implantation endometrium of pregnant mice which suggested that Activin A and Inhibinαcollaboratively were involved in the process of implantation.6.The number ofimplantation sites was greatly decreased by the intrauterine injection with Activin A polyclonal antibody in the early morning of the day 4 of pregnancy. |
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