PROSTAGLANDIN E2 RECEPTOR IN SALIVARY GLANDS

唾液腺中的前列腺素 E2 受体

• 批准号: 2131184
• 负责人: CHI-YING WU-WANG
• 金额: $ 3.99万
• 依托单位: UNIV OF MED/DENT OF NJ-NJ MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-04-01 至 1996-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2131184
• 关键词: ADP ribosylation; SDS polyacrylamide gel electrophoresis; acinar cell; adenylate cyclase; affinity chromatography; autoradiography; biological signal transduction; cell membrane; cholera toxin; cyclic AMP; enzyme activity; gel filtration chromatography; guanosine triphosphate; laboratory rat; phosphorylation; prostaglandin E; prostaglandin receptor; protein purification; receptor binding; sublingual gland; submandibular gland

Prostaglandins (PGs) are potent biologically active compounds, known to act as local tissue hormones capable of triggering a multiplicity of physiological responses. In salivary submandibular and sublingual glands, PGs, especially those of the E series, have been demonstrated to regulate salivary flow and mucin secretion, and may play a role in the control of electrolyte concentrations in saliva. However, the cellular mechanism of PGEs action in salivary glands is poorly understood. The PI has recently characterized a specific PGE2 receptor in crude membranes of salivary submandibular glands. The binding of PGE2 to a specific receptor on cell membrane may be the initial step leading to its biological actions within the cells. To better elucidate the role of PGE2 receptor in salivary glands at the cellular level, it is important to purify, identify and characterize the receptor protein, as well as to investigate the intracellular signal transduction pathway mediated by PGE2 and its receptor. The specific aims of this proposed project are: Aim #1. Purification, identification and characterization of PGE2 receptor in acinar cells of salivary glands. Acinar cells from submandibular and sublingual glands will be isolated. Various subcellular membrane fractions will be prepared and analyzed or PGE2 receptor binding. The receptor-enriched subcellular fraction(s) will be used for receptor purification and identification. Aim #2. Study of PGE2 receptor-mediated signal transduction in acinar cells of salivary glands. Semipurified acinar plasma membrane will be prepared and used to study the receptor-coupled intracellular second messenger system, which involves the functional associations among PGE2, receptor and N protein in the adenylate cyclase system. In situ protein phosphorylation after receptor binding will also be examined. A temporal correlation between the in situ protein phosphorylation and the intracellular cAMP production under the influence of PGE2 in a dose- dependent manner will be investigated. Purification and identification of receptor protein in salivary gland acini will not only support the data of biochemical features of the receptor, but also be a necessary prerequisite for future receptor studies at both the cellular and molecular levels. The proposed study will also reveal important information regarding the intracellular processing of signal after the receptor binding. The Principal Investigator (PI) anticipates that this proposed research will pave the avenues towards a better understanding of the regulatory role played by PGE2 and its receptor in salivary glands, and expects to elucidate the mechanism underlie the links between the intracellular signals and the biological functions triggered by PGE2 in salivary glands.

前列腺素 (PG) 是有效的生物活性化合物，已知 作为局部组织激素能够触发多种 生理反应。在颌下腺和舌下腺的唾液中， PG，尤其是 E 系列的 PG，已被证明可以调节 唾液流量和粘蛋白分泌，并可能在控制中发挥作用 唾液中的电解质浓度。然而，细胞机制 PGEs 在唾液腺中的作用尚不清楚。 PI 最近 表征了唾液粗膜中的特定 PGE2 受体 颌下腺。 PGE2与细胞上特定受体的结合 膜可能是导致其生物作用的第一步 细胞。更好地阐明PGE2受体在唾液中的作用 腺体在细胞水平上，纯化、识别和 表征受体蛋白，并研究 PGE2及其介导的细胞内信号转导通路 受体。该拟议项目的具体目标是： 目标#1。 PGE2 受体的纯化、鉴定和表征 存在于唾液腺的腺泡细胞中。 腺泡细胞来自颌下和 舌下腺将被分离。 各种亚细胞膜 将制备并分析PGE2受体结合的级分。这 富含受体的亚细胞部分将用于受体 纯化和鉴定。 目标#2。 PGE2受体介导的腺泡信号转导研究 唾液腺细胞。半纯化的腺泡质膜将 制备并用于研究受体偶联的细胞内第二 信使系统，涉及 PGE2 之间的功能关联， 腺苷酸环化酶系统中的受体和N蛋白。原位蛋白 受体结合后的磷酸化也将被检查。一个暂时的 蛋白质原位磷酸化与 在一定剂量的 PGE2 的影响下，细胞内 cAMP 的产生 依赖方式将受到调查。 唾液腺受体蛋白的纯化及鉴定 acini 不仅支持生化特征数据 受体，也是未来受体研究的必要前提 在细胞和分子水平上。拟议的研究还将 揭示有关细胞内加工的重要信息 受体结合后发出信号。首席研究员 (PI) 预计这项拟议的研究将为 更好地了解 PGE2 及其作用 唾液腺中的受体，并期望阐明其机制 之间的联系的基础 PGE2触发的细胞内信号和生物学功能 在唾液腺中。