HORMONAL CONTROL OF OVIDUCTAL SECRETORY PROTEINS

我们的目标是增加我们对输卵管成分的了解 分泌物以及它们可能如何影响建立和维持 通过描述未受精者之间的分子相互作用来描述怀孕 可能由输卵管介导的卵子、精子和受精卵 蛋白质。这项提案的最初目标是描述 猪输卵管合成和分泌的蛋白质 雌激素和孕激素的影响；也就是对 为确定怀孕提供适当的环境。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 目的：研究输卵管蛋白合成和分泌的调节。 内源性类固醇在发情周期和早孕期间，并通过 卵巢切除动物外源性应用类固醇。这将是 在外植体培养中使用标记的前体完成的假设 保持体内状态的分泌活动特征 在受控的体外环境中。放射性标记的蛋白质将是 2D-PAGE后经荧光照相鉴定。不同职能部门的作用 还将评估合成和分泌中的片段(A、I、AIJ)。 另一个目标将是研究卵巢介导的超微结构变化 类固醇的光镜和电子显微镜观察 卵巢切除动物，并与分泌特异性激素的变化相关 蛋白质。一个主要的目标将是纯化主要的输卵管分泌物 蛋白质，特别是当卵子在雌激素支配下产生的蛋白质 精子也是存在的。生化特征将包括 测定MR、PI、碳水化合物含量和任何酶的活性。 利用这些纯化的蛋白，将获得多克隆的单特异性抗血清 为确定合成部位的免疫细胞化学研究做好准备， 颗粒形成和胞外分布。这将允许更多 对激素控制的精确研究。另一个目标是评估 这些蛋白质具有免疫抑制或补体抑制活性， 对妊娠的建立和维持很重要。这项研究将 在不同的动物模型中提供重要数据：1)卵巢类固醇 输卵管分泌蛋白的调控，2)鉴定 雌激素和孕激素特异性蛋白，3)生化 这些蛋白的特性，以及4)卵巢类固醇诱导的 超微结构改变。

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.