REGULATION OF PROSTAGLANDIN RECEPTORS IN MYOMETRIUM

描述（由申请人提供）：前列腺素（PG），由 子宫内组织似乎在子宫内膜异位症的发生和/或 维持分娩。子宫肌层对前列腺素的反应和敏感性 可能受多种PG受体的差异表达所控制， G蛋白偶联受体超家族的一部分， 跨膜结构域。对任何特定的前列腺素的反应 (e.g. PGE2）可能取决于受体亚型及其亚型， 任何特定的时间。目前还没有关于表达的全面数据。 妊娠子宫肌层PG受体及其激素调节。是 目前还表明，怀孕的人类子宫可能显示出功能性的 足月时的区域化，由此下节松弛以允许通过 胎儿的头部，但上段收缩，以排出胎儿。这 区域化可能通过PG受体的差异表达介导 同种型。这些受体的克隆和测序提供了机会 研究存在的亚型和亚型， 并随着分娩和荷尔蒙控制而变化。总体假设 有待测试的是，子宫对肾上腺素的反应性部分是 由收缩和舒张PG受体的差异表达调节 在子宫肌层中存在同种型，并且这种表达部分受到子宫内膜的调节， 雌激素/孕激素本研究的目的是（1）研究mRNA和 收缩性FPI、EP 1、EP 3和TP的蛋白质表达和定位， 妊娠大鼠子宫肌层（第16天至第1天）的舒张性IP、EP 2和EP 4受体 在妊娠动物中，通过激素调控来改变 研究期间内分泌（雌激素/孕激素）环境（第16天以后）（2） 收缩性和舒张性PG受体的表达和定位 获得配对的上和下段妊娠人子宫肌层样品， 无论是足月还是早产，来自正在分娩或未分娩的患者（3）， 用共聚焦显微镜研究PG受体在子宫肌层的细胞定位 在组织切片和分离的肌细胞上进行显微镜检查（4），以确定 在EP2基因的5 '侧翼区的反应元件，其决定 在术语表达上的变化。这些研究将提供全面的 哪些受体在子宫肌层中表达， 随着妊娠和分娩开始的变化以及 雌激素/孕激素对这种表达的影响。人类研究还将揭示， 在人类子宫肌层中存在功能区室化， PG受体的差异表达。共焦显微镜将提供 受体的细胞定位和启动子分析信息 将揭示控制子宫内膜癌基因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.