CHANGING MYOMETRIAL PHENOTYPES DURING PREGNANCY

怀孕期间子宫肌层表型的变化

• 批准号: 6435884
• 负责人: PAUL C. MACDONALD
• 金额: $ 19.72万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6435884
• 关键词: G protein; adenylate cyclase; biopsy; birth; cyclic AMP; estrogen receptors; gestational age; guanylate cyclase; hormone regulation /control mechanism; human tissue; myometrium; northern blottings; phenotype; phospholipase C; pregnancy; protein kinase C; radioimmunoassay; tissue /cell culture; transfection; western blottings

The research proposed is designed to characterize the functional phenotypes of the uterus during pregnancy (before and during labor) and to define the cellular changes that establish the phenotypic differences. The studies described are closely aligned with those designed to determine the biomolecular mechanisms by which the transitions in parturition phase are effected. The studies are based on the hypothesis that thre are three major components of a fail-safe system that ensures myometrial quiescence for the first 90-95 percent of human pregnancy: (i) the actions of estrogen-progesterone; (ii) hormones, neuropeptides and eicosanoids acting via heptahelical receptors linked to Galpha/S adenylyl cyclase; and, (iii) natriuretic peptide-activated membrane receptor/guanylyl cyclase, which effects an increase in intracellular cGMP. The 3 system components that serve to effect myometrial quiescence must be muted late in pregnancy as uterine phase O is suspended. There also is a group of ligands that act in myometrium via heptahelical receptors linked to Galpha/i/q/11-causing inhibition of adenylyl cyclase and activation of phospholipase C (PLC). An increase in responsiveness to Galpha/i/q/11-linked receptor ligands likely facilitates the establishment of uterine phase 1, ultimately eventuating in the completion of the uterine transition to phase 2, i.e., active labor. Myometrial membrane preparations from pre-term and term pregnancies (before and during labor) will be used to evaluate responsiveness to multiple (9 different) ligands operative via Galpha/S- linked heptahelical receptors that activate adenylyl cyclase. The cAMP- activation state of the myometrium will be evaluated by determining endogenous levels of cAMP and the steady-state activity of protein kinase A (PKA). Various components of the system will be explored to define the nature of cellular changes that account for phenotypic differences among tissues: heptahelical receptors and linkage to G-proteins, total (activatable) adenylyl cyclase, G-protein expression and subcellular distribution. Responsiveness of myometrial membrane preparation to activation of membrane receptor/guanylyl cyclase by natriuretic peptides will be evaluated together with the levels of endogenous cGMP and the steady state cGMP-induced protein kinase. Tissue responsiveness to ligands that act via Galpha/i/q/11,-linked heptahelical receptors to activate phospholipase C also will be assessed. Estrogen receptor-transfected human myometrial cells will be used to define the cause of cellular changes that account for the changes in functional phenotypes.

所提出的研究旨在表征功能 怀孕期间（临产前和临产期间）子宫的表型 定义建立表型差异的细胞变化。这 所描述的研究与旨在确定 分娩阶段转变的生物分子机制 生效了。这些研究基于三个假设： 确保子宫肌层静止的故障安全系统的主要组成部分 对于人类怀孕的前 90-95%： (i) 雌激素-孕激素； (ii) 激素、神经肽和类二十烷酸的作用 通过与 Galpha/S 腺苷酸环化酶连接的七螺旋受体；以及，（三） 利钠肽激活膜受体/鸟苷酸环化酶， 影响细胞内 cGMP 的增加。 3 个系统组件 影响子宫肌层静止的作用必须在妊娠后期减弱，因为 子宫O期暂停。还有一组配体发挥作用 子宫肌层通过七螺旋受体与 Galpha/i/q/11 相关 抑制腺苷酸环化酶和激活磷脂酶 C (PLC)。一个 对 Galpha/i/q/11 连接受体配体的反应性增加 最终可能促进子宫第一阶段的建立 最终完成子宫向第二阶段的过渡，即 积极劳动。早产儿和足月子宫肌膜制剂 怀孕（分娩前和分娩期间）将用于评估 对通过 Galpha/S- 起作用的多个（9 个不同）配体的响应 连接的七螺旋受体激活腺苷酸环化酶。 cAMP- 子宫肌层的激活状态将通过确定来评估 内源性 cAMP 水平和蛋白激酶的稳态活性 A（PKA）。将探索系统的各个组件来定义 细胞变化的本质导致了表型差异 组织：七螺旋受体和与 G 蛋白的连接，总 （可激活）腺苷酸环化酶、G 蛋白表达和亚细胞 分配。子宫肌膜制备物的反应性 利钠肽激活膜受体/鸟苷酸环化酶 将与内源性 cGMP 水平和 稳态 cGMP 诱导的蛋白激酶。组织对配体的反应性 通过 Galpha/i/q/11 连接的七螺旋受体起作用以激活 磷脂酶 C 也将被评估。雌激素受体转染的人 子宫肌细胞将用于确定细胞变化的原因 解释功能表型的变化。