The Role of Trp Proteins in Myometrial Calcium Dynamics

Trp 蛋白在子宫肌层钙动态中的作用

• 批准号: 7066668
• 负责人: BARBARA M SANBORN
• 金额: $ 18.51万
• 依托单位: COLORADO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7066668
• 关键词: calcium channel; calcium flux; calcium indicator; calcium transporting ATPase; clinical research; confocal scanning microscopy; estrogens; female; hormone regulation /control mechanism; human subject; laboratory rat; muscle contraction; myometrium; polymerase chain reaction; pregnancy; protein protein interaction; tissue /cell culture; voltage gated channel; western blottings

DESCRIPTION (provided by applicant): Understanding the mechanisms regulating myometrial intracellular free Ca (Cai) dynamics is critical to understanding the control of myometrial activity. Signal-regulated calcium entry (also called capacitative entry) is broadly defined here to include both receptor- and store-operated entry. This pathway has been implicated in regulation of membrane potential and tone in smooth muscle and may play a role in the regulation of myometrial contractile activity by hormones, growth factors and cytokines. Hence regulated calcium entry may participate in control of uterine contractile activity during pregnancy and parturition. We found that rat and human myometrial cells respond to more than one signal-regulated calcium stimulus and express a number of TrpC mRNAs and proteins implicated in this type of calcium entry. The overall goal of this study is to understand the role of TrpC proteins in the regulation of myometrial Cai dynamics and control of contractile activity. Aim 1 will determine the relative importance of signal-regulated Ca entry in the control of myometrial Cai dynamics, measured by fluorescence imaging, and to contractile activity in uterine strips. Aim 2 will delineate the contribution of specific TrpC proteins to signal-regulated calcium entry and Cai dynamics in human myometrial cells. The effects of both overexpression and suppression of TrpC expression on myometrial cell Cai dynamics will be determined and linked to specific responses. Aim 3 will examine the regulation of TrpC expression and of signal-regulated Ca entry by other signaling pathways in myometrium. These studies will utilize Q-RT-PCR, immunoblot and Ca imaging approaches in human myometrial cells and the pregnant rat model. Aim 4 will examine the influence of TrpC subtype expression on TrpC association with myometrial plasma membrane and possible effects of signal-regulated Ca entry on activity of other membrane proteins. These studies will use scanning confocal microscopy and subcellular fractionation approaches. These studies promise new insights into the role of this tightly regulated calcium entry mechanism in the control of myometrial Cai dynamics and myometrial contraction and may lead to design of new approaches for controlling myometrial activity during pregnancy and labor.

描述（由申请人提供）：了解子宫内膜胞内游离钙（Cai）动力学的调节机制对于理解子宫内膜活动的控制至关重要。信号调节的钙离子进入（也称为容性进入）在这里被广泛定义为包括受体和储存操作的钙离子进入。该通路与平滑肌膜电位和张力的调节有关，并可能通过激素、生长因子和细胞因子调节子宫肌收缩活动。因此，调节钙的进入可能参与控制妊娠和分娩期间子宫收缩活动。我们发现，大鼠和人类肌层细胞对不止一种信号调节的钙刺激做出反应，并表达许多与这种钙进入有关的TrpC mrna和蛋白质。本研究的总体目标是了解TrpC蛋白在肌内膜动力学调节和收缩活性控制中的作用。