ROLE OF ADENYL PURINES IN MYOMETRIAL FUNCTION

该项目提出，腺基嘌呤，一种已知能引起 子宫收缩，参与人体的调节 分娩。这项研究的具体目的是发现 来源、结合部位和分子信号事件 腺苷对子宫肌条的收缩作用。这个 工作假说是腺基嘌呤，包括腺苷和来自 一种或多种来源刺激子宫肌层平滑的嘌呤能受体 肌肉导致子宫收缩。我们建议腺基嘌呤 参与或开始分娩，并因此可以提供一个焦点 来了解人类的早产。研究将在以下情况下进行 组织和新鲜分离的细胞 人类。研究计划将分为三个领域： 调查。(1)我们将确定腺基嘌呤在 子宫。高效液相色谱和放射性标记研究将调查 可能的嘌呤能神经及其对嘌呤的共同释放 子宫肌层的肾上腺素能和胆碱能神经。旁分泌释放 来自子宫肌层本身的平滑肌或子宫内膜的嘌呤 也将接受测试。(2)腺嘌呤的受体靶点将是 在细胞和组织中进行放射性配基结合研究 将决定嘌呤能受体的存在和分布 环行肌和纵行肌的P1和P2亚型。这些 还将使用方法来确定监管的性质 受体对激动剂的亲和力和偶联蛋白的作用 GTP结合蛋白在这些受体分子中的作用。 药理学研究将确定腺基嘌呤的作用 在怀孕的子宫里。组织的机械收缩， 细胞离子电流和电流的电生理测量 将进行受体作用的生化测量。生化和 电生理学研究将使用组织片段和纯化 并将集中于嘌呤在血管内皮细胞中的作用。 细胞内钙离子的调节。生化研究将包括测量 肌醇磷酸、花生四烯酸释放、环核苷酸 代谢和蛋白激酶的作用。细胞成像使用 将使用荧光pH染料来确定对 磷脂酶A2通过刺激N+/H+交换。细胞内 研究细胞和组织中的钙波动将使用 细胞内的荧光染料。

This project proposes that adenyl purines, compounds known to cause contraction of uterus, participate in the regulation of human parturition. The specific aims of this research are to discover the origins, binding sites and molecular signaling events that subserve the contractile action of adenyl purines in uterine smooth muscle. The working hypothesis is that adenyl purines, both adenosine and ATP from one or more sources stimulate purinergic receptors in myometrial smooth muscle leading to uterine contraction. We propose that adenyl purines participate in, or initiate labor and as such, may provide a focal point for understanding preterm labor in humans. Studies will be conducted with tissues and freshly isolated cells from both pregnant guinea pigs and humans. The research plan will be separated into three areas of investigation. (1) We will determine the origins of adenyl purines in the uterus. HPLC and radiolabeling studies will investigate the role of putative purinergic nerves as well as the co-release of purines from adrenergic and cholinergic nerves in myometrium. Paracrine release of purines from the smooth muscle of myometrium itself, or from endometrium will also be tested. (2) The receptor targets for adenyl purines will be investigated with radioligand binding studies in cells and tissues and will determine the presence and distribution of purinergic receptors of the P1 and P2 subtype in circular and longitudinal smooth muscles. These methods will also be used to determine the nature of regulation of receptor affinity for agonist and the role of coupling proteins such as GTP-binding proteins in the molecular action of these receptors.(3) Pharmacological studies will characterize the actions of adenyl purines in pregnant uterus. Mechanical contraction of tissues, electrophysiological measurements of cellular ionic currents and biochemical measurements of receptor action will be made. Biochemical and electrophysiological studies will employ tissue segments and purified smooth muscle cells and will concentrate on the role of purines in the regulation of cellular Ca+2. Biochemical studies will include measurement of inositol phosphates, arachidonic acid release, cyclic nucleotide metabolism and the action of protein kinases. Cellular imaging using fluorescent pH dyes will be used to determine the regulation of phospholipase A2 through stimulation of N+/H+ exchange. Intracellular calcium fluctuations will be studied in cells and tissues using intracellular fluorescent dyes.