GROWTH FACTOR CONTROL OF APOPTOSIS DURING PLACENTATION

植入期间细胞凋亡的生长因子控制

• 批准号: 2202158
• 负责人: BRUCE C MOULTON
• 金额: $ 16.4万
• 依托单位: UNIVERSITY OF CINCINNATI
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 1997-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2202158
• 关键词: antisense nucleic acid; apoptosis; calcium flux; cell differentiation; cell growth regulation; decidua; embryo implantation; endometrium; estrogens; gel electrophoresis; gene expression; growth factor receptors; hormone regulation /control mechanism; immunocytochemistry; in situ hybridization; laboratory rat; northern blottings; progesterone; receptor binding; transforming growth factors; western blottings

Cell death by apoptosis takes place during the growth and development of vertebrate tissues and during the adult function of tissues. As a complement to mitosis, apoptosis has a vital function in the uterus in maintaining the homeostasis of cell number during the estrous cycle and in the tissue remodeling inherent in blastocyst implantation and development of the placenta. The proposed research will examine the pattern of expression and hormonal control of apoptosis in endometrial epithelial and stromal cells during blastocyst implantation and during decidualization. Our hypothesis is that progesterone and estrogen secretion during early pregnancy controls not only the growth and differentiation of endometrial cells but also controls their regression by apoptosis through the control of the synthesis of TGF-beta's and/or their receptors. To examine this hypothesis, the following specific aims are proposed: (l) We will determine the control of apoptosis by progesterone and estrogen and establish the kinetics of this cellular process in luminal epithelial cells in response to a deciduogenic stimulus and in stroma cells during decidualization in hormone-treated ovariectomized rats, pseudopregnant rats, and pregnant rats. Antiprogestins will be used in these studies to examine progesterone control of endometrial transformation following implantation. (2) We will determine the growth factor control of stromal cell apoptosis focusing upon TGF-beta2 as the growth factor which controls stromal apoptosis during decidual regression by a paracrine and/or autocrine mechanism. The synthesis of TGF-beta's the specific cells will be examined by immunohistochemistry and in situ hybridization. We will measure receptor levels for TGF-beta's and characterize the receptor proteins. (3) The biochemical mechanisms by which TGF-beta initiates apoptosis in endometrial stromal cells will be examined. We will measure the response to TGF-beta of specific genes which have been identified as early participants in the initiation of apoptosis. The role of these genes in stromal apoptosis will be determined by inhibiting their expression with antisense oligodeoxynucleotides corresponding to translation start regions. The hypothesis that an influx of Ca++ is required for apoptosis will be tested by examining the effect of TGF-beta on intracellular levels of Ca++ of cultured stromal cells and by determining effects of blocking Ca++ influx on apoptosis. Completion of these studies should contribute to our understanding of fundamental cellular and biochemical mechanisms of blastocyst implantation and decidualization.

细胞凋亡导致的细胞死亡发生在细胞的生长和发育过程中， 脊椎动物组织和组织的成年功能期间。作为 作为有丝分裂的补充，细胞凋亡在子宫中具有重要的功能， 维持细胞数量的稳态在发情周期和 胚泡植入和发育中固有的组织重塑 胎盘拟议的研究将审查模式， 子宫内膜上皮细胞凋亡的表达和激素调控， 基质细胞在胚泡植入和蜕膜化过程中。 我们的假设是，孕激素和雌激素分泌在早期 妊娠不仅控制子宫内膜的生长和分化， 细胞，而且还通过控制细胞凋亡来控制它们的退化。 TGF-β和/或其受体的合成。审查这个 假设，提出了以下具体目标：（l）我们将 确定孕酮和雌激素对细胞凋亡的控制， 在管腔上皮细胞中建立这种细胞过程的动力学 细胞对蜕膜刺激的反应，以及基质细胞在 卵巢切除大鼠子宫内膜蜕膜化，假孕 老鼠和怀孕的老鼠这些研究中将使用抗孕激素， 检查孕酮对子宫内膜转化的控制， 置入(2)我们将确定基质细胞的生长因子控制 细胞凋亡集中在TGF-β 2作为生长因子， 旁分泌和/或 自分泌机制特定细胞合成TGF-β 免疫组化和原位杂交检测。我们将 测量TGF-β的受体水平并表征受体 proteins. (3)转化生长因子β启动的生化机制 将检测子宫内膜基质细胞中的凋亡。我们将测量 对特定基因的TGF-β的反应，这些基因已被鉴定为 细胞凋亡启动的早期参与者。这些基因的作用 在基质细胞凋亡中的作用将通过抑制它们的表达来确定 具有对应于翻译起始的反义寡脱氧核苷酸 地区细胞凋亡需要Ca++内流的假说 将通过检查TGF-β对细胞内水平的影响来测试 的Ca++的培养基质细胞，并通过确定的影响， Ca++内流对细胞凋亡的影响。 完成这些研究将有助于 对于我们理解基本的细胞和生化机制 胚泡着床和蜕膜化。