REGULATION OF CONNEXIN-43 GENE EXPRESSION IN UTERUS

这项研究计划描述了旨在揭示 连接蛋白43基因在子宫肌层中的调控机制。 该基因编码形成细胞-细胞的缝隙连接蛋白。 频道。这些通道负责电信号的传播 通过肌肉的脉动来实现子宫的同步收缩 分娩过程中的肌肉。与心肌不同，连接蛋白43是 在任何时候都存在，如果它是按宪法表达的， 子宫肌层细胞-细胞通道只有在分娩开始时才会出现 分娩后不久就消失了。这种细胞-细胞通道的诱导 形成受雌激素和黄体酮的控制。建议数 连接蛋白43基因调控的研究将在大鼠连接蛋白43上进行 包含6.7 kb上游调控序列的基因组克隆。它 包括6个半回文雌激素和3个黄体酮反应元件。 转录蛋白与这些和其他调节因子的结合 元素将通过DNA足迹分析和迁移率变化进行研究 化验。启动子研究将使用包含以下内容的构建体进行 启动子的不同片段，包括上游调控序列， 与荧光素酶的报告基因相关联。这些构造将是 在转基因小鼠中表达，不同激素的影响将是 下定决心。单个启动子元素的贡献将是 通过诱变分析进行评估。最后，反式作用转录 因子将从一个cdna表达文库中分离出来并进行鉴定。 这项研究的长期目标是了解 子宫肌层间隙连接蛋白43基因表达的调控 怀孕和分娩时。这项研究具有临床意义，因为 已有研究表明，早产与早产有关 子宫肌层缝隙连接的出现。一旦不同的人相互作用 含有连接蛋白43基因调控元件的激素效果更好 了解，更好的治疗和预防早产的养生法 劳动可能是设计出来的。

This research proposal describes experiments designed to reveal the mechanism by which the connexin43 gene is regulated in the myometrium. This gene codes for the gap junction protein that forms cell-cell channels. These channels are responsible for the propagation of electrical pulses through the muscle to allow synchronous contraction of the uterine muscle during labor. In contrast to heart muscle, where connexin43 is present at all times and were it is expressed constitutively, the myometrial cell-cell channels appear only at the onset of labor and disappear shortly after parturition. This induction of cell-cell channel formation is under the control of estrogen and progesterone. The proposed study of connexin43 gene regulation will be done with a rat connexin43 genomic clone that contains 6.7 kb of upstream regulatory sequence. It includes 6 half-palindromic estrogen and 3 progesterone response elements. The binding of transcriptional proteins to these and other regulatory elements will be studied by DNA footprinting analysis and mobility shift assays. Promoter studies will be performed with constructs that contain various segments of the promoter, including upstream regulatory sequences, linked to the reporter gene for luciferase. These constructs will be expressed in transgenic mice, and the effect of different hormones will be determined. The contribution of individual promoter elements will be assessed by mutagenic analysis. Finally, trans-acting transcription factors will be isolated from a cDNA expression library and characterized. The long-term goal of this research is to gain an understanding of the regulation of connexin43 gene expression in the myometrium during pregnancy and at parturition. This research is clinically relevant because it has been shown that premature labor is associated with the premature appearance of myometrial gap junctions. Once the interaction of different hormones with the regulatory elements of the connexin43 gene is better understood, better regimens for the treatment and prevention of premature labor may be designed.