Regulation of cell survival in the pregnant rat endometrium.

怀孕大鼠子宫内膜细胞存活的调节。

• 批准号: 238501-2013
• 负责人: Asselin, Eric
• 金额: $ 2.4万
• 依托单位: Université du Québec à Trois-Rivières
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2015
• 资助国家: 加拿大
• 起止时间: 2015-01-01 至 2016-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=570434
• 关键词: Regulation; cell; survival; pregnant; rat

Recognition and maintenance of pregnancy are complex processes involving both maternal and embryonic signals. In the rodents during early implantation, endometrial epithelial cells undergo apoptosis. Later, the stromal cells undergo decidualization, which is followed by decidua basalis (DB) regression, involving apoptosis of decidual cells. The failure to establish such a communication has been shown to be the most important cause of infertility among mammals. However, the molecular mechanisms governing endometrial cell apoptosis are not yet defined. Interestingly, our laboratory has showed that individual TGF-b isoforms (TGF-b1, -b2 and -b3) are differently expressed in the uterus during rat pregnancy: TGF-b1 and TGF-b2 are present during implantation and DB regression, whereas TGF-b3 isoform is only present during DB regression. We have also provided evidence that distinct TGF-b can activate distinct signaling pathways in endometrial cells. This strongly suggests that TGF-b can play an important role depending of the situation during pregnancy and we hypothesize that sex steroids progesterone and 17b-estradiol, the hormones of pregnancy, might also be involved in this process. The overall and long term objective of the proposed research program is to examine and understand the regulation of expression as well as the endocrine, paracrine and autocrine activity of the TGF-b isoforms in the endometrium during the two important phases of pregnancy: 1) embryo implantation and 2) DB regression. Epithelial and stromal cells as well as whole rat uteri from different Days of pregnancy, pseudopregnat and estrous cycle will be used for in vivo and in vitro studies. Experimental approaches will include molecular and biochemical techniques, upregulation with expression vectors and down-regulation of key proteins using shRNA, biochemical in vivo inhibition of signaling pathways, apoptosis in situ and by flow cytometry and immunofluorescence. Information gained from these studies will help to understand the importance of TGF-b isoforms during these critical reproductive situations and will provide important clues for successful establishment and maintenance of pregnancy.

妊娠的识别和维持是涉及母体和胚胎信号的复杂过程。在啮齿类动物的早期着床过程中，子宫内膜上皮细胞会发生凋亡。随后，基质细胞经历蜕膜化，随后发生基底蜕膜（DB）退化，涉及蜕膜细胞的凋亡。未能建立这种通讯已被证明是哺乳动物不育的最重要原因。然而，控制子宫内膜细胞凋亡的分子机制尚未明确。有趣的是，我们的实验室表明，在大鼠怀孕期间，各个TGF-b亚型（TGF-b1、-b2和-b3）在子宫中表达不同：TGF-b1和TGF-b2在着床和DB回归期间存在，而TGF-b3亚型仅在DB回归期间存在。我们还提供了证据表明不同的 TGF-b 可以激活子宫内膜细胞中不同的信号通路。这强烈表明 TGF-b 可以根据怀孕期间的情况发挥重要作用，我们假设性类固醇黄体酮和 17b-雌二醇（怀孕激素）也可能参与这一过程。拟议研究计划的总体和长期目标是检查和了解妊娠两个重要阶段期间子宫内膜中 TGF-b 同种型的表达调节以及内分泌、旁分泌和自分泌活性：1) 胚胎着床和 2) DB 退化。妊娠、假妊娠和动情周期不同天数的上皮细胞和基质细胞以及整个大鼠子宫将用于体内和体外研究。实验方法将包括分子和生化技术、使用shRNA上调表达载体和下调关键蛋白、信号通路的体内生化抑制、原位细胞凋亡以及流式细胞术和免疫荧光。从这些研究中获得的信息将有助于了解 TGF-b 同工型在这些关键生殖情况下的重要性，并将为成功建立和维持妊娠提供重要线索。