REGULATION OF PROSTAGLANDIN RECEPTORS IN MYOMETRIUM

描述(申请人提供)：前列腺素(PG‘s)由 子宫内组织似乎在启动和/或 维持分娩。子宫肌层对PG的反应和敏感性 可能由不同PG受体的差异表达所控制，这些受体是 G蛋白偶联受体超家族的一部分，它有七个 跨膜跨域。对任何特定前列腺素的反应 (例如PGE2)可能取决于受体亚型及其亚型 任何特定的时间。目前还没有关于基因表达的全面数据。 妊娠子宫肌层PG受体及其激素调节。它是 目前还提示，怀孕的人类子宫可能会表现出功能性 地区化的条件，即较低的部分放松以允许通过 胎头，但上段收缩以驱逐胎儿。这 PG受体的差异表达可能介导区域化 异构体。这些受体的克隆和测序提供了机会 研究目前存在的异构体和亚型，以及它们在整个妊娠过程中的个体发育 并随着分娩和荷尔蒙的调节而改变。总体假设 需要检验的是，子宫对前列腺素的反应性在一定程度上 收缩和松弛PG受体的差异表达调控 子宫肌层中的异构体，这种表达在一定程度上受激素调节 通过雌激素/黄体酮。本研究的目的是(1)研究信使核糖核酸和 收缩因子FPI、EP1、EP3和TP的蛋白表达与定位 妊娠16~1日龄大鼠子宫肌层IP、EP2和EP4受体的松弛作用 产后一天)，以及在怀孕动物中通过荷尔蒙操作来改变 足月(第16天起)内分泌(雌激素/孕酮)环境(2)研究 心肌收缩和松弛PG受体的表达和定位 配对获取上、下节段妊娠人子宫肌层标本 无论是在足月还是早产的患者(3) 用激光共聚焦技术研究PG受体在子宫肌层的细胞定位 组织切片和分离的心肌细胞的显微镜观察(4)以确定 决定EP2基因5‘侧翼区的反应元件 在学期中表达的变化。这些研究将提供一个全面的 图中哪些受体在子宫肌层中表达，如何表达 随妊娠和临产时间的变化及其影响 雌激素/孕激素对这种表达的影响。人体研究还将揭示如果 在人的子宫肌层中有一种功能分区，与 PG受体的差异表达。共聚焦显微镜将提供 受体的细胞定位信息和启动子分析 将揭示控制子宫表达的潜在调控位点 分娩时的受体。

DESCRIPTION (provided by applicant): Prostaglandin's (PG's) produced by intrauterine tissues appear to have a major role in the initiation and/or maintenance of parturition. The response and sensitivity of myometrium to PG's may be governed by differential expression of various PG receptors which are part of the superfamily of G protein coupled receptors that have seven transmembrane spanning domains. The response to any particular prostaglandin (e.g. PGE2) may depend on the receptor isoforms and their subtypes expressed at any particular time. Currently there is no comprehensive data on expression of PG receptors in the pregnant myometrium or their hormonal regulation. It is also currently suggested that the pregnant human uterus may show a functional regionalization at term whereby the lower segment relaxes to allow passage of the fetal head, but the upper segment contracts to expel the fetus. This regionalization may be mediated via differential expression of PG receptor isoforms. The cloning and sequencing of these receptors offers the opportunity to study isoforms and subtypes present, their ontongeny throughout gestation and changes with parturition and hormonal manipulation. The overall hypothesis to be tested is that uterine responsivity to prostaglandins is in part regulated by differential expression of contractile and relaxatory PG receptor isoforms in myometrium and that this expression is hormonally regulated in part by estrogen/progesterone. The objectives of the study are (1) to study mRNA and protein expression and localization for contractile FPI, EP1, EP3, and TP and relaxatory IP, EP2, and EP4 receptors in pregnant rat myometrium (day 16 to one day postpartum) and in pregnant animals with hormonal manipulation to alter the endocrine (estrogen/progesterone) milieu at term (day 16 onwards) (2) to study expression and localization of contractile and relaxatory PG receptors in paired upper and lower segment pregnant human myometrial samples obtained either at term or preterm from patients who are or are not in labor (3) to study the cellular localization of PG receptors in myometrium using confocal microscopy on tissue sections and isolated myocytes (4) to determine the response elements in the 5' flanking region of the EP2 gene that determine alterations in expression at term. These studies will provide a comprehensive picture of which receptors are expressed in myometrium, how the expression changes with gestation and the onset of labor and the effect of estrogen/progesterone on this expression. The human studies will also reveal if there is a functional compartmentalization in human myometrium associated with differential expression of PG receptors. The confocal microscopy will provide information on cellular localization of receptors and the promoter analyses will reveal potential regulatory sites for control of expression of uterine receptors at parturition.