HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS

输卵管分泌蛋白的激素控制

• 批准号: 3448230
• 负责人: WILLIAM C BUHI
• 金额: $ 4.67万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-04-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3448230
• 关键词: antibody; early embryonic stage; egg /ovum; electron microscopy; estrogens; estrus; fallopian tubes; fertilization; gel electrophoresis; gel filtration chromatography; genital secretion; hormone regulation /control mechanism; immunochemistry; ovariectomy; progesterone; sperm capacitation

Our goals are to increase our knowledge of the composition of oviductal secretions and how they may affect the establishment and maintenance of pregnancy by characterizing molecular interactions between the unfertilized ovum, spermatozoa and fertilized ovum which may be mediated by oviductal proteins. The initial objective of this proposal is to characterize the proteins synthesized and secreted by the porcine oviduct under the influence of estrogen and progesterone; that is, proteins which contribute to the appropriate environment for establishment of pregnancy. This includes maintenance of the ovum, capacitation of spermatozoa, fertilization and early embryonic development. Two of our major aims will be to study regulation of synthesis and secretion of oviductal proteins by endogenous steroids during the estrous cycle and early pregnancy, and by exogenously administered steroids in ovariectomized animals. This will be accomplished using labelled precursors in explant cultures assuming secretory activities characteristic of the in vivo situation is maintained in a controlled in vitro setting. Radiolabelled proteins will be identified by fluorography after 2D-PAGE. The role of different functional segments (A,I,AIJ) in synthesis and secretion will also be assessed. Another aim will be to study ultrastructural changes mediated by ovarian steroids with light and electron microscopy in both intact and ovariectomized animals, and correlate changes with secretion of specific proteins. A major aim will be to purify the major oviductal secretory proteins, specifically those produced under estrogen dominance when ovum and sperm are present. Biochemical characterization will include determination of Mr, pI, carbohydrate content, and any enzyme activity. Using these purified proteins, polyclonal monospecific antiserum will be prepared for immunocytochemical studies to define the site of synthesis, granule formation and extracellular distribution. This will allow more precise studies of hormone control. Another aim will be to assess whether these proteins possess immunosuppressive or complement inhibitory activity, important for establishment and maintenance of pregnancy. This study will provide important data in a different animal model on: 1) ovarian steroid regulation of oviductal secretory proteins, 2) identification of estrogen-specific and progesterone-specific proteins, 3) biochemical characterization of these proteins, and 4) ovarian steroid-induced ultrastructural changes.

我们的目标是增加我们对输卵管的组成的了解， 分泌物，以及它们如何影响建立和维持 通过表征未受精的 卵子、精子和受精卵，可能通过输卵管介导 proteins. 本提案的初步目标是描述 猪输卵管合成和分泌的蛋白质 雌激素和孕激素的影响;也就是说，蛋白质， 适合怀孕的环境。 这 包括卵子的维持，精子的获能， 受精和早期胚胎发育。 我们的两个主要目标将 研究输卵管蛋白质合成和分泌的调控， 内源性类固醇在发情周期和早期妊娠，并通过 在切除卵巢的动物中外源性给予类固醇。 这将是 在外植体培养物中使用标记的前体完成， 保持体内情况的分泌活性特征 在受控的体外环境中。 放射性标记的蛋白质将被 2D-PAGE后通过荧光法鉴定。 不同职能的作用 还将评估合成和分泌中的片段（A，I，AIJ）。 另一个目的将是研究卵巢癌介导的超微结构变化， 类固醇与光学和电子显微镜在两个完整的， 卵巢切除动物，并与特定的分泌相关的变化 proteins. 一个主要的目的将是纯化主要的输卵管分泌 蛋白质，特别是那些在雌激素主导下产生的蛋白质， 和精子的存在。 生化表征将包括 Mr、pI、碳水化合物含量和任何酶活性的测定。 使用这些纯化的蛋白质，将制备多克隆单特异性抗血清。 制备用于免疫细胞化学研究以确定合成位点， 颗粒形成和细胞外分布。 这将允许更多 激素控制的精确研究。 另一个目标是评估是否 这些蛋白质具有免疫抑制或补体抑制活性， 对怀孕的建立和维持很重要。本研究将 在不同的动物模型中提供了重要的数据：1）卵巢类固醇 输卵管分泌蛋白的调节，2）鉴定 雌激素特异性蛋白和雌甾酮特异性蛋白，3）生物化学 这些蛋白质的特征，和4）卵巢类固醇诱导的 超微结构变化