STRUCTURAL AND FUNCTIONAL CHARACTERIZATION OF PLACENTAL LACTOGENS

胎盘泌乳素的结构和功能特征

• 批准号: 6301811
• 负责人: FRANK J. TALAMANTES
• 金额: $ 20.96万
• 依托单位: UNIVERSITY OF CALIFORNIA SANTA CRUZ
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2001-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6301811
• 关键词: cell differentiation; chemical binding; epidermal growth factor; female; hormone binding protein; hormone regulation /control mechanism; in situ hybridization; insulinlike growth factor; interleukin 6; laboratory mouse; liver cells; pregnancy; pregnancy circulation; prolactin; protein purification; protein structure function; somatomammotropin; transforming growth factors; tumor necrosis factor alpha

The mouse placenta produces two placental lactogens (PLs); mPL-I and mPL- II. The long term objectives of the project are to understand the functions of these hormones in various target tissues and to understand what regulates their production and concentrations in the fetal and maternal blood. Although relatively little is known about the biological activities and regulation of secretion of PLs in various species, it is clear that PLs participate in several important processes during pregnancy. Investigating the physiology of PLs will contribute to a better understanding of the processes that influence fetal health and survival; during pregnancy. These studies are a continuation of previous work in this laboratory on the biological and regulation of secretion of mPL-I and mPL-II. Several experiments include examining the regulation of secretion of proliferin (PLF) and PLF-related protein (PRP), which are proteins that are structurally similar to the PLs and are also produced by the mouse placenta. The specific aims of this project are: (1) The primary structure of calcyclin, a stimulator of mPL-II secretion will be determined. The protein will be localized in the conceptus by in situ hybridization, and the presence of binding sites for decidual calcyclin in the placenta will be assessed. Effects of decidual calcyclin on PL-I, PLF and PRP secretion will be examined in vivo. (2) Effects of epidermal growth factor, transforming growth factors alpha and beta, tumor necrosis factor-alpha, interleukin (IL)-beta, and IL-6 alone and in combinations, will be determined on giant cell differentiation and on mPL-I, mPL-II, PLF and PRP secretion by placental cells from various days of pregnancy. The sites of production of these agents in the conceptus will be determined by in situ hybridization. (3) The structure of the genes for mPL-I and mPL-II will be determined, and the promoter region will be examined for the presence of known response elements. Putative regulators of mPL-I and mPL-II expression identified in this way will be examined in vitro and in vivo for effects on mPL-I and mPL-II secretion. (4) A putative stimulator of mPL-I secretion produced by the pituitary gland will be characterized and if it appears to be novel, it will be purified and characterized. (5) An inhibitor of prolactin secretion produced by the placenta will be purified. Its interaction with mPL-I and mPL-II in regulating prolactin secretion will be examined in vitro. (6) A novel 29K insulin-like growth factor binding protein (IGBP) whose production is induced by mPLs will be purified and its primary structure will be determined. The liver will be examined for the presence of 29K IGFBP. The gestational profile of 29K IGFBP in liver and mammary gland will be determined. Regulation of 29K IGFBP by mPL-I and mPL-II in liver and mammary gland will be examined. (7) Proteins regulated by mPL-I and mPL-II in maternal liver will be identified.

小鼠胎盘产生两种胎盘催乳素（PL）; mPL-I和mPL-I。 二. 该项目的长期目标是了解 这些激素在各种靶组织中的功能，并了解 是什么调节了它们在胎儿体内的产生和浓度， 母亲的血 尽管对生物学的了解相对较少， 活性和调节PL的分泌在各种物种中，它是 明确PL参与了几个重要的过程， 怀孕 研究PL的生理学将有助于 更好地了解影响胎儿健康的过程， 生存;怀孕期间。 这些研究是以前研究的延续。 在这个实验室工作的生物和调节分泌的 mPL-I和mPL-II。 几项实验包括检查监管 增殖素（PLF）和PLF相关蛋白（PRP）的分泌， 与PL结构相似的蛋白质， 被老鼠胎盘所吸引 本项目的具体目标是：（1） 一级结构的钙周期蛋白，刺激mPL-II分泌将是 测定 蛋白质将通过原位定位在孕体中 杂交和蜕膜钙周期蛋白结合位点的存在 将对胎盘进行评估。 蜕膜钙周期蛋白对磷脂酶I、 将在体内检查PLF和PRP分泌。 (2)表皮的影响 生长因子，转化生长因子α和β，肿瘤坏死 因子-α、白细胞介素（IL）-β和IL-6的单独和组合， 将根据巨细胞分化和mPL-I，mPL-II， 不同妊娠天数胎盘细胞的PLF和PRP分泌。 这些药物在孕体中的产生部位将是 通过原位杂交测定。 (3)基因的结构 将测定mPL-I和mPL-II，并将测定启动子区。 检查已知响应元素的存在。 推定 以这种方式鉴定的mPL-I和mPL-II表达的调节因子将被 在体外和体内检测对mPL-I和mPL-II分泌的影响。 (4)垂体产生的mPL-I分泌的假定刺激物 腺将被表征，如果它看起来是新的，它将是 纯化和表征。(5)一种催乳素分泌抑制剂 胎盘产生的毒素会被净化 它与mPL-I的相互作用 和mPL-II在调节催乳素分泌中的作用将在体外进行检查。 (6)一种新的29 K胰岛素样生长因子结合蛋白（IGBP），其 mPL诱导的产生将被纯化，其一级结构 将被确定。 将检查肝脏中是否存在29 K IGFBP。 29 K胰岛素样生长因子结合蛋白在肝脏和乳腺中的妊娠分布 将被确定。 mPL-I和mPL-II对肝脏29 K IGFBP的调节作用 检查乳腺。 (7)受mPL-I调节的蛋白质和 将鉴定母体肝脏中的mPL-II。