HORMONAL CONTROL OF DELAYED IMPLANTATION

延迟着床的荷尔蒙控制

• 批准号: 3310514
• 负责人: RODNEY A MEAD
• 金额: $ 8.57万
• 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/3310514
• 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来确定机制： 光周期，下丘脑，垂体和 卵巢相互作用控制停止和重新启动 胚胎发育和植入的物种表现出 延迟着床。 该提案特别寻求 提供更多的证据来支持语料库 鼬科食肉动物如家养雪貂的黄体， 斑点臭鼬分泌一种蛋白质化合物， 用于诱导胚泡植入。 我建议进一步 纯化和化学表征雪貂黄体植入 因子（LIF），并确定哪些营养因子刺激其 体外合成和分泌。 然后我提议证明 通过对雪貂和斑点臭鼬的被动免疫， LIF的抗体将抑制植入。 另一个实验 旨在确定怀孕的哪几天或 假孕LIF在雪貂中分泌。 我亦建议 进一步验证LIF仅由雪貂的CL产生，并 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/或胚泡， 提供证据表明存在受体位点， 化合物. 这将通过研究特异性结合来实现 （125 I）-LIF对子宫和胚泡质膜的粘附。 进一步 将通过以下方法证明特异性受体位点： 提取交联的（125 I）-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的具体影响 通过测量cAMP的变化， 蛋白质、RNA和DNA的合成。