REGULATION OF PROSTAGLANDIN RECEPTORS IN MYOMETRIUM

前列腺素(PG S)似乎在启动过程中起着重要作用 和/或维持分娩。前列腺素的抑制作用 合成被用作早产宫缩的一种机制。 并不是没有对胎儿产生潜在的有害副作用。这个 子宫肌层对PG的反应和敏感性可能受 不同PG受体的差异表达，其中许多已被 最近进行了克隆和测序。PG受体是 具有七个跨膜的G蛋白偶联受体超家族 跨域。每种前列腺素都有一个特定的受体 异构体连接到不同的信号转导途径 收缩或松弛反应。受体的交替剪接 信使核糖核酸转录本产生亚型亚型，这些亚型可与 不同的信号转导途径具有不同的作用。因此， 对任何特定前列腺素(如PGE2)的反应可能取决于 受体亚型及其亚型在任何特定时间表达。 存在于子宫肌层中的受体亚型以前 以药理手段为特点的。最新的克隆和测序 这些受体提供了批判性研究异构体的机会 以及亚型的存在，它们在整个妊娠过程中的个体发育和变化 分娩和荷尔蒙控制。不幸的是，具体来说 目前还没有针对受体亚型的抗体和探针。一个 这项建议的主要目标是开发试剂以 促进这些研究。需要检验的总体假设是 子宫对前列腺素的反应性在一定程度上受 收缩和松弛PG受体的差异表达 子宫肌层中的异构体，这种表达受激素调节 通过雌激素/黄体酮。 这项研究的三个目标是(1)识别和综合 每个受体亚型的独特多肽序列可提高抗体水平 用于免疫组织化学和免疫印迹研究(2)分离cdna S精选大鼠和人子宫肌层受体异构体使用 原位杂交和Northern分析；(3)研究 米非司酮对妊娠大鼠子宫肌层受体亚型的调节 调控雌激素和孕激素的合成和作用。 PG受体的发生、个体发育及其调控的研究进展 在子宫肌层是开始探索其用途的关键第一步 刺激抑制特定受体作为一种新的选择性 发酵术的方法。必要的第一步是发展 有助于这项研究和进一步用于探测的高质量试剂 受体结构/功能关系。

Prostaglandins (PG s) appear to have a major role in the initiation and/or maintenance of parturition. Inhibition of prostaglandin synthesis is used as a mechanism of tocolysis in preterm labor, though not without potentially deleterious side effects for the fetus. The response and sensitivity of myometrium to PG's may be governed by differential expression of various PG receptors, many of which have been recently cloned and sequenced. The PG receptors are part of the superfamily of G protein-coupled receptors that have seven transmembrane spanning domains. Each prostaglandin species has a specific receptor isoform linked to a distinct signal transduction pathway giving contraction or relaxatory responses. Alternate splicing of receptor mRNA transcripts give rise to isoform subtypes that can be linked to different signal transduction pathways with diverse actions. Hence, the response to any particular prostaglandin (e.g. PGE2) may depend on the receptor isoforms and their subtypes expressed at any particular time. The receptor isoforms present in myometrium have previously been characterized by pharmacologic means. The recent cloning and sequencing of these receptors offers the opportunity to critically study isoforms and subtypes present, their ontogeny throughout gestation and changes with parturition and hormonal manipulation. Unfortunately, specific antibodies and probes to the receptor isoforms are not available. A principal objective of this proposal is to develop reagents to facilitate these studies. 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 by estrogen/progesterone. The three objectives of the study are to (1) identify and synthesize unique peptide sequences for each receptor isoform to raise antibodies for immunohistochemistry and western blotting studies (2) isolate cDNA s for selected receptor isoforms from rat and human, myometrium for use in in situ hybridization and northern analysis and (3) study the regulation of receptor isoforms in pregnant rat myometrium by manipulation of estrogen and progesterone synthesis and action. Description of the occurrence, ontogeny and regulation of PG receptors in myometrium is a crucial first step in beginning to explore the use of stimulation of inhibition of specific receptors as a novel selective means of tocolysis. A necessary first step is the development of quality reagents to aid in this study and for further use in probing receptor structure/function relationships.