HORMONAL CONTROL OF DELAYED IMPLANTATION

延迟着床的荷尔蒙控制

• 批准号: 3310516
• 负责人: RODNEY A MEAD
• 金额: $ 12.56万
• 依托单位: UNIVERSITY OF IDAHO
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-06-01 至 1992-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3310516
• 关键词: antibody; corpus luteum; cyclic AMP; early embryonic stage; embryo /fetus; embryo implantation; ferrets; growth /development; high performance liquid chromatography; hormone receptor; hormone regulation /control mechanism; immunocytochemistry; mammalian embryology; pituitary gonadal axis; protein biosynthesis; radiotracer; uterus

The long term goal of this research is to define the mechanisms by which photoperiod, hormones of the hypothalamus, pituitary and ovary interact in controlling cessation and reinitiation of embryonic development and implantation in a species exhibiting obligate delay of implantation. This proposal specifically seeks to Provide additional evidence to support the theory that corpora lutea of mustelid carnivores such as the domesticated ferret and spotted skunk secrete a proteinacious compound that is essential for induction of blastocyst implantation. I propose to further purify and chemically characterize the ferret luteal implantation factor (LIF) and to determine which trophic factor(s) stimulate its synthesis and secretion in vitro. I then propose to demonstrate by passive immunization of ferrets and spotted skunks, that antibodies to LIF will inhibit implantation. Another experiment is designed to determine on which days of pregnancy or pseudopregnancy LIF is secreted in ferrets. I also propose to further verify that LIF is only produced by CL of ferrets and to explore its subcellular distribution and mode of secretion using immunocytochemistry. Another series of experiments is designed to determine whether LIF acts on the uterus and or blastocysts and to provide evidence for the presence of receptor sites to this compound. This will be accomplished by studying specific binding of (125I)-LIF to uterine and blastocyst plasma membranes. Further demonstration of specific receptor sites will be accomplished by extraction of cross-linked (125I)-LIF-receptor complexes, followed by electrophoresis and autoradiography. The last set of experiments is designed to determine the specific effects of LIF on its target organs by measuring such things as changes in cAMP, protein, RNA and DNA synthesis in the uterus and or blastocysts.

这项研究的长期目标是通过以下方式定义机制： 光周期、下丘脑、垂体和 卵巢在控制停止和重新启动中相互作用 物种中的胚胎发育和植入 强制延迟着床。 该提案特别寻求 提供额外的证据来支持语料库的理论 鼬类食肉动物的黄体，例如驯化的雪貂和 斑点臭鼬分泌一种必需的蛋白质化合物 用于诱导囊胚植入。 我建议进一步 雪貂黄体植入的纯化和化学表征 因子（LIF）并确定哪些营养因子刺激其 体外合成和分泌。 然后我建议展示 通过对雪貂和斑点臭鼬进行被动免疫， LIF 抗体会抑制植入。 另一个实验 旨在确定怀孕的哪几天或 假妊娠 雪貂会分泌 LIF。 我也提议 进一步验证LIF仅由雪貂的CL产生并 使用以下方法探索其亚细胞分布和分泌方式 免疫细胞化学。 另一系列实验旨在 确定 LIF 是否作用于子宫和/或囊胚，并 为该受体位点的存在提供证据 化合物。 这将通过研究特异性结合来完成 (125I)-LIF 进入子宫和囊胚质膜。 更远 特定受体位点的演示将通过以下方式完成 提取交联的 (125I)-LIF-受体复合物，然后 通过电泳和放射自显影。 最后一组 实验旨在确定 LIF 的具体效果 通过测量 cAMP 的变化等来影响其靶器官， 子宫和/或囊胚中的蛋白质、RNA 和 DNA 合成。