Assessment of alpha4, alphav, beta1 and beta3 integrins expression throughout the implantation window phase in endometrium of a mouse model of polycystic ovarian syndromes

Endometrial integrin expression changes might be a reason for implantation failure in polycystic ovarian syndromes [PCOS]. Assessment of integrin genes and proteins expression upon endometrium in the PCOS experimental mouse model was the main goal of this study. 30 NMRI female mice were equally divided into control, experimental [PCOS; received estradiol valerate [40 mg/kg]] and sham group [received; olive oil]. After 8 weeks, each group was hyper stimulated by 7 IU PMSG and then, after 48hrs, 7 IU HCG was injected. Vaginal plaque was checked. After 5 days, Progesterone and estradiol levels and endometrial tissues were investigated to evaluate of alpha4, alphav, beta1 and beta3 integrins gene and protein by qPCR method and immunohistochemistry, respectively. Tissue samples were assessed and showed that level of progesterone was significantly decreased in PCOS group. Results of molecular part in the amount of alphav, beta3, beta1 and alpha4 gene expressions showed a great difference in beta3 and alphav genes expressions between experimental groups, alphav, beta3, alpha4 and beta1 proteins in the endometrial stroma in the control group were expressed, but they were not detected in PCOS group. According to the results, integrins had different expression patterns in different areas of the endometrium; such as epithelial and stromal. It seems that in PCOS, this pattern has changed and the results might have a great influence on implantation failure. Therefore, this study suggests that a great attention to this problem may be essential in patients who are involved

Expression of alpha4, alphav, beta1 and beta3 integrins during the implantation window on blastocyst of a mouse model of polycystic ovarian syndromes

It has been hypothesized that blastocyst integrin expression changes can affect the spontaneous miscarriage in polycystic ovarian syndromes [PCOS]. In this study, the profile of integrin genes and proteins was investigated on blastocyst of the PCOS experimental mouse model. 30 NMRI female mice were equally divided into 3 groups: control, experimental [PCOS that was injected estradiol valerate [40 mg/kg]]. After 8 weeks, each group was hyper stimulated by PMSG and HCG. Vaginal plaque was checked, and mice were investigated 5 days after the test. Progesterone and estradiol levels were determined; alpha4, alpha v, beta 1 and beta 3 integrin genes and protein of blastocysts were examined by real time PCR method and immunohistochemistry, respectively. Estradiol level was significantly increased [p

Effects of selenium, calcium and calcium ionophore on human oocytes in vitro maturation in a chemically defined medium

In vitro maturation [IVM] of human oocytes is an emerging procedure quickly incorporated into the world of assisted reproductive technologies. As an effective method of in vitro maturation, several studies have reported the critical role of differentions on activating the complex process involved in both gamete maturation and fertilization. In this study, we supplemented a chemically defined medium with different combinations of selenium, calcium and calcium ionophore concentrations to obtain the best rate of human oocytes maturation, survival, and fertilization. As an experimental study, Three combinations of [selenium [5 microg/ml], calcium [5 microg/ml] and calcium ionophore [1 microg/ml]], [selenium [10 microg/ml], calcium [7 microg/ml] and calcium ionophore [2 microg/ml]] and [selenium [15 microg/ml], calcium [10 microg/ml] and calcium ionophore [5 microg/ml]] added to the chemically defined medium and the morphology of oocytes assessed after 22-24 hours in vitro maturation of the oocytes. The highest percentage of MII [meiosis II] oocytes [68%], developing beyond the morula [20.1%] and the blastocyst formation [11.1%] observed in oocytes treated with 15microg/ml selenium, 10microg/ml calcium and 5microg/ml calcium ionophore. Moreover, we showed the significant rate of survival in each three combinations after 36, 72 and 96 hours. Maturation and activation of oocytes may be triggered by changes in intracellular ion concentrations as second messengers in signal transduction pathways. Here, we received the highest percentage of in vitro maturation and fertilization among three combinations of selenium, calcium and calcium ionophore treatments. Using this combination of ions beside other factors might be useful for the enrichment of the human oocytes IVM medium

correlation between the endometrial integrins and osteopontin expression with pinopodes development in ovariectomized mice in response to exogenous steroids hormones

The ovariectomized animals are good models to evaluate the effect of different steroid hormone treatments on implantation events and the pattern of integrin expression. Therefore, this study was performed to compare the expression of integrins and osteopontin [OPN] in correlation with pinopode development in ovariectomized mice endometrium which was subjected to steroid hormones. Ovariectomized mice were subjected to estrogen, progesterone and estrogen-progesterone hormones. Their uterine horns were evaluated for integrin expression by immunohistochemistry and real-time RT-PCR and for pinopode development by transmission and scanning electron microscopic studies. No immunostaining for integrin and OPN molecules were detected in the endometrium of non-ovariectomized mice except in metestrus phase. The alpha4 and beta 1 integrin genes were expressed in all phases of estrous cycle. In ovariectomized mice, no reaction was detected in the endometrium of control, sham and estrogen-treated groups, but in both progesterone-treated groups, all examined genes were expressed. There was not any correlation between pinopodes and integrin expression in ovariectomized mice. The progesterone is more effective on endometrial integrin expression than estrogen and differences in the expression pattern of integrins reflect their important and different roles in embryo implantation. The pinopodes may have minor effect in mice implantation or have some delay in their expressions in ovariectomized mice which were subjected to exogenous hormones

[ changes of alpha v, beta 3, alpha 4, beta 1 integrins expression and osteopontin their ligands in murine estrous cycle]

Considering the importance of integrin molecules in the implantation and lack of sufficient information in the expression pattern of these molecules in various phases of estrous cycle. It seemed to be necessary to investigate these molecules in mouse endometrial during the various phases of oestrous cycle. Female NMRI mice [n=15] aged 6-8 weeks were studied. Various phases of estrous cycle including: proestrus, estrus, metestrus and diestrus were determined by vaginal smear. The mice were sacrificed [at least 3 per each phase] by cervical dislocation and the tissues were obtained from the middle 1/3 part of their uterine horns at each phase then the cryosections at thicknesses between 8-10 micro were obtained. Then the immunohistochemistry were done for integrins of alpha4, beta1, alphav, beta3 and their ligand osteopontine. The integrins were expressed only in the metestrous phase of oestrous cycle in the different locations of mouse endometrium. The positive reactions were observed for alphav, alpha4 and beta3 in the apical and basal membrane of glandular epithelium. Also the positive reaction for beta1 was found in surface and glandular epithelium as well as stroma. The osteopontin expression was seen in the apical membranes of surface and glandular epithelium and was not seen in other locations. It seems that expression of integrins in endometrium is based on their role in the implantation, therefore the molecules alpha4, beta1 and OPN that are expressed on the surface epithelial may be involve in the adhesion of cell to cell and integrins of alphav, beta3 that are expressed in the glandular