| The uterus is an important reproductive organ. It is necessary for many of life’s pivotal cellular events and biological processes, such as embryo implantation, decidualization, placentation and labor. Therefore, it has become increasingly critical to explore and clarity how uterine functions are completed and potential regulatory factors for these events. Recently, there is increasing evidence that bone morphogenetic protein (BMP) family and nitric oxide (NO) play vital roles in the uterine functions and development, such as Bmp2 has an important role in the decidualization, and NO could contribute to the local control of myometrial contractility. In this regard, the present study is designed to investigate the expression of bone morphogenetic protein 2/4 (Bmp2/4) and related components of the BMP signaling pathway in mice uterus as well as the cellular expression and immunolocalization of nitric oxide synthase (NOS) isoforms and soluble guanylyl cyclase (sGC) subunits in postnatal porcine uteri using a kind of animal models. The mainly findings and conclusions are as follows.1 Dynamic expression of Bmp2/4 and related components of the BMP signaling pathway in mice uterus during postnatal developmentThe present study is designed to investigate the expression of bone morphogenetic protein2/4 (Bmp2/4) and related components of the BMP signaling pathway in mice uterus during postnatal development using real-time polymerase chain reaction (PCR). Uterine samples were collected from ICR mice aged 1,3,10,15,21 and 29 days and dissected free of surrounding tissue. Our results showed that the expression level of Bmp2 had a gradually increasing tendency and reached its highest value at postnatal days 21 and 29, whereas Bmp4 exhibited a slight oscillatory pattern during the postnatal periods, specifically, the expression level of Bmp4 was significantly higher at postnatal days 15 and 21 than other periods (P<0.05). There was statistically significant difference between the expression levels of Bmp2 and that of Bmp4 (P<0.05). The relative quantity of Bmprla was significantly higher at postnatal days 3 and 29 than that at other periods investigated (P<0.05). The expression levels of Bmprlb and Bmpr2 had a gradually increasing tendency during the postnatal periods. Moreover, the level of Bmprlb mRNA was significantly lower compared with that of Bmprla and Bmpr2 mRNA at the corresponding stages (P<0.05). All three R-Smads were differentially expressed in the mouse uterus and the expression levels of Smadl and Smad5 were significantly higher compared with that of Smad8 (P<0.05). In addition, the relative abundance of Smad4 exhibited significant fluctuations. Our findings strongly suggested that Bmp2/4 signaling pathway might play key roles in the regulation of organization and stratification of endometrial stroma and the maintenance of uterine endometrium in mice uterus during postnatal development.2 Expression of Bmp2/4 and related components of the BMP signaling pathway in the mouse uterus during the estrus cycleThe objective of the present study was to investigate the roles of bone morphogenetic protein family members in the mouse uterus during the estrus cycle by real-time polymerase chain reaction (PCR) and immunohistochemistry. Real-time PCR analysis showed that the expression level of Bmp2 was significantly higher at proestrus than that at estrus and metestrus (P<0.05). The relative abundance of Bmp4 exhibited significant fluctuations. There were no statistically significant difference between the expression level of Bmp2 and that of Bmp4. The expression levels of Bmprla and Bmpr2 remained unchanged during estrus cycles. However, the level of Bmprlb mRNA significantly decreased at estrus (P<0.05), increasing subsequently at metestrus (P<0.05). Furthermore, the level of Bmprlb mRNA was significantly lower compared with that of Bmpr1a and Bmpr2 mRNA in the corresponding stages (P<0.05). All three R-Smads were differentially expressed in the mouse uterus and the expression level of Smad1 and Smad5 were significantly higher compared with that of Smad8 (P<0.05). In addition, the expression level of Smad4 did not substantially change throughout the estrus cycle. Immunohistochemical experiments revealed BMP2 protein was differentially and mainly localized in the uterine luminal and glandular epithelial cells throughout the estrus cycle. The results demonstrated differential expressions and localizations of bone morphogenetic protein family members in the mouse uterus, which supported the hypothesis that BMP signaling was differentially involved in the control of the degradation and remodeling of mouse endometrium during the estrus cycle. These findings will be helpful in exploring the overall mechanisms regulating cycling changes of mouse uterus.3 Expression of Bmp2/4 and related components of the BMP signaling pathway in the uterus of pregnant miceThe present study is designed to investigate the roles of ovarian steroids and Bmp2/4 signaling pathway during pregnancy in mice by Radioimmunoassay (RIA) and real-time polymerase chain reaction (PCR). The results from RIA showed that concentration of E2 was reached its lowest level on days 5 and 7, whereas it was not significantly change in any of other periods investigated. Concentration of P4 had a gradual increase tendency during early and mid pregnancy, but it had a gradual reduction tendency until D18. The P/E value showed gradually increased during the early pregnancy whereas it reached the minimum before delivery. The results from real-time PCR revealed that the expression level of Bmp2 had a gradual tendency during early pregnancy, and reached its highest value on day 7, then sharply decreased on day 12 (P<0.05). In contrast, the relative abundance of Bmp4 exhibited a slight oscillatory pattern. Specifically, the expression level of Bmp4 was significantly higher on PP3 than that at other stages of pregnancy (P<0.05). There was statistically significant difference between the expression levels of Bmp2 and that of Bmp4 (P<0.05). The relative abundance of Bmprla had a gradual reduction tendency from day 1 to day 12 whereas it showed gradual upward trend in the following days. The relative quantity of Bmpr1b was significantly higher on day 3 than that at other periods of implantation (P<0.05). Furthermore, the relative quantity of Bmprlb was not changed significantly from days 12 to PP1, but increased significantly on PP3 (P<0.05), compared with other stages of pregnancy. Bmpr2 showed extremely stable expressed before delivery, whereas it significantly increased after delivery (P<0.05). In addition, the level of Bmprlb mRNA was significantly lower compared with that of Bmprla and Bmpr2 mRNA in the corresponding stages (P<0.05). R-Smads were differentially expressed in the mouse uterus and the expression levels of Smadl and Smad5 were significantly higher compared with that of Smad8 (P<0.05). Furthermore, the relative abundance of Smad4 mRNA remained unchanged throughout pregnancy, whereas it was significantly increased on PP3 (P<0.05). These results strongly suggested that ovarian steroids and Bmp2/4 signaling pathway might play key roles in the regulation of establishment and maintenance of pregnancy and parturition.4 Roles and ovarian steroids regulation of BMP2/4 signaling pathway in the mouse uterus during the peri-implantation periodsThe present study is designed to investigate the temporal expression patterns of Bmp2/4 and related components of the BMP signaling pathway during peri-implantation period (days 1-8) and its regulation in the uterus by ovarian steroid hormones or/and their antagonists. The results showed that expression level of Bmp2 had a gradual increase tendency and reached its highest value on day 7. The relative abundance of Bmp4 showed gradual upward trend from day 1 to 3, whereas it had a gradual reduction tendency in the following days. There was statistically significant difference between the expression levels of Bmp2 and that of Bmp4 (P<0.05). The expression patterns of Bmprs were largely similar to that of Bmp4 except that relative quantity of Bmprla and Bmpr2 mRNA remained high on day 4. Moreover, the level of Bmprlb mRNA was significantly lower compared with that of Bmprla and Bmpr2 mRNA in the corresponding stages (P<0.05). All three R-Smads were differentially expressed in the mouse uterus and the expression levels of Smadl and Smad5 were significantly higher compared with that of Smad8 (P<0.05). The relative abundance of Smad4 was significantly higher from day 2 to 5 than that at other days (P<0.05). In addition, injection of E2 induced body weight gain for the ovariectomized mice but this phenomenon was not observed in response to other treatments. The treatment with combination of P4 and E2 up-regulated the expression level of Bmp4 mRNA. Our findings suggested that Bmp2/4 signaling pathway might play a critical role in murine decidualization during the peri-implantation, and ovarian steroids are potential regulator of BMP family members.5 Cell-specific expression and immunolocalization of nitric oxide synthase isoforms (NOS) and soluble guanylyl cyclase (sGC) a and P subunits in postnatal porcine uteriThe aim of the present study was to investigate the cellular expression and immunolocalization of nitric oxide synthase (NOS) isoforms and soluble guanylyl cyclase (sGC) subunits in postnatal porcine uteri. Immunohistochemical experiments showed that three isoforms of NOS were mainly localized in the uterine luminal and glandular epithelium and myometrium, and the intensity of immunostaining for iNOS and eNOS was increased gradually with temporal development of the postnatal uterus. In addition, sGC subunits, sGCal and (3, were present in the uterine luminal and glandular epithelium, myometrium and stromal cells. The uterine NOS activity data showed that the total NOS and iNOS activity were significantly increased at postnatal days 21 (P<0.05) and 35 (P<0.05). Although constitutive NOS activity was increased at postnatal day 21, it decreased subsequently at postnatal day 35 (P<0.05). Immunoblot analysis revealed that iNOS protein expression was significantly increased at postnatal days 21 (P<0.05) and 35 (P<0.05). Furthermore, sGCα1 protein expression was not significantly changed throughout days 7 to 35. Collectively, our findings suggest that NO/cGMP signaling is involved in the process of postnatal porcine uterine development.In a word, we used a kind of animal models to investigate the expression of Bmp2/4 and related components of the BMP signaling pathway in mice uterus as well as the cellular expression and immunolocalization of NOS isoforms and sGC subunits in postnatal porcine uteri using real time PCR, IHC and wester blot. Our findings suggested that Bmp2/4 signaling pathway appear to have instrumental roles in controlling cellular events and biological processes involved in the regulation of endometrial adenogenesis, degradation and remodeling of endometrium, implantation, decidualization and pregnancy through their action associated with cell proliferation, differentiation and apoptosis. In addition, endometrium- and myometrium-derived NO contributes to the local control of inhibition of myometrial contractions and thereby causes uterine relaxation in the postnatal pig. |
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