INITIATION OF LABOR: MYOMETRIAL PROTEIN SYNTHESIS

分娩开始：子宫肌蛋白合成

• 批准号: 3315715
• 负责人: WILLIAM B CURRIE
• 金额: $ 12.49万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-12-01 至 1987-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3315715
• 关键词: antibody; antibody formation; autoradiography; birth; clone cells; creatine kinase; electrofocusing; estrogens; fluorescence microscopy; gel electrophoresis; gel filtration chromatography; gene expression; genetic markers; genetic transcription; genetic translation; hormone regulation /control mechanism; immunoelectrophoresis; membrane activity; muscle contraction; myometrium; organ culture; progestins; radioimmunoassay; radiotracer; reagent /indicator; sarcolemma; smooth muscle; ultracentrifugation

The actions of regulatory steroids are manifested in myometrium through altered synthesis of proteins. Protein components of the sarcolemma such as agonist receptors, ion channels and gap junctions are affected. Major changes in excitability and electrocoupling of the smooth muscle cells are the basis for the functional transition from the quiescent organ of pregnancy to the active and expulsive organ of labor. Interference with these changes may provide a means of preventing pre-term labor. Research should be focussed on determining where potential therapeutic agents may act using in vitro methods and animal studies. Our research will identify proteins, the synthesis of which is subject to regulation by estrogens and progestins, that underly some functional changes initiating labor. Creatine Kinase-BB (formerly Induced Protein) will serve as a model protein to test our hypothesis that progesterone serves mainly to tonically constrain synthesis of "activator" proteins. These proteins are expected to be of two main classes; a) those that are destined to become incorporated into the membrane with roles closely linked to the process of excitation such as agonist rceptors, junctional components and ion channels; and b) enzymes capable of modifying intrinsic sarcolemmal components to render the cell more excitable. Methods are in place to enable in vitro steroid manipulations on surviving explants and myometrial smooth muscle cells have been established in cloned culture. These techniques will permit the exposure of additional sarcolemmal and soluble proteins subject to tight steroidal control by affinity labelling or pulse-labelling of surface and newly synthesized proteins in cloned smooth muscle cells, followed by 2D electrophoresis. Immunoreagents prepared against these steroid regulated proteins will be evaluated for their suitability as probes for studying dynamic changes in the membrane of myometrial cells being exposed to steroidal manipulations. Methods will be developed using the rabbit as a model species because of clear-cut steroidal control of the myometrium; it is planned to extend the work later by applying the methodology to primates with the eventual goal of studying human myometrial cells established into cell culture.

调节性类固醇的作用表现在子宫肌层， 改变蛋白质的合成。 肌膜的蛋白质成分， 作为激动剂受体，离子通道和缝隙连接受到影响。 主要 平滑肌细胞的兴奋性和电偶联的变化是 从静止器官的功能转变的基础 怀孕到分娩的主动和排出器官。 干扰 这些变化可以提供一种预防早产的方法。 研究 应该集中在确定潜在的治疗剂可以 使用体外方法和动物研究。 我们的研究将确定 蛋白质，其合成受雌激素调节， 孕激素是引发分娩的某些功能变化的基础。 肌酸激酶-BB（以前的诱导蛋白）将作为模型蛋白 为了验证我们的假设，黄体酮主要是 抑制“激活剂”蛋白的合成。 预计这些蛋白质 有两个主要类别; a）那些注定要成为 被纳入膜中，其作用与细胞分裂过程密切相关 兴奋，如激动剂受体，连接组件和离子 通道;和B）能够修饰内在肌膜的酶 使细胞更容易兴奋。 方法已到位， 能够在存活的外植体和子宫肌层上进行体外类固醇操作 已经在克隆培养物中建立了平滑肌细胞。 这些 技术将允许暴露额外的肌膜和可溶性 通过亲和标记进行严格类固醇控制的蛋白质，或 脉冲标记克隆的光滑表面和新合成的蛋白质 肌肉细胞，然后进行2D电泳。 制备的免疫试剂 针对这些类固醇调节的蛋白质，将评估它们的 适合作为研究膜动态变化的探针， 子宫肌层细胞暴露于类固醇操作。 方法将 由于明确的， 子宫肌层的类固醇控制;计划以后扩展这项工作 通过对灵长类动物应用这种方法， 将人子宫肌层细胞建立到细胞培养物中。