ACTH/MSH PEPTIDES & PHORBOL ESTERS ON PKC PHOSPHORYLATION & CA LEVELS IN SHR

ACTH/MSH 肽

• 批准号: 6107728
• 负责人: CLEO A HUGHES
• 金额: $ 7.78万
• 依托单位: MORGAN STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-03-01 至 2000-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6107728
• 关键词: adrenocorticotropic hormone; biological signal transduction; calcium; enzyme activity; hormone regulation /control mechanism; hypertension; immunoprecipitation; isozymes; laboratory rat; melanocyte stimulating hormone; mesencephalon; pathologic process; phorbols; phosphatidylinositols; phosphorylation; prosencephalon; protein kinase C; rhombencephalon; spontaneous hypertensive rat; synaptosomes; western blottings

Description (Adapted from Application): Phosphoinositide signaling system and calcium have been reported to play a role in the regulation of blood pressure. In preliminary studies, it has been established by immunoblot and immunohistochemical studies that PKC isozymes alpha, beta, gamma, delta, epsilon, and zeta are expressed in synaptosomes isolated from brain regions of normal and hypertensive rats. It has also been established that gamma-2-MSH alters the expression of PKC isozymes as well as PKC activity in hypertensive rats. It has been reported that ACTH-like peptides derived either from the extracellular space affect the B-50 (GAP-43) phosphoprotein through interaction with the presynaptic membrane of the nerve terminal. Also, the brain is rick in PKC and Ca2+ and it is sensitive to hypertension, which causes a variety of vascular and neuronal cerebral changes. Taken together, it is hypothesized that in hypertension, there is a link between peptide (i.e., gamma-2-MSH, ACTH[4-10], and ACTH] PKC-mediated phosphorylation of specific PKC isozymes, which results in changes in PKC-mediated phosphorylation of specific proteins as well as alterations in the levels of calcium. In order to understand the role of the central nervous system and the phosphoinositol signaling pathway in hypertension, the investigators plan to use: (1) protein phosphorylation, immunoprecipitation, and Western immunoblot assays to identify the specific proteins that are phosphorylated as a result of the activation of PKC-delta, -epsilon, and-gamma in the forebrain, midbrain, and hindbrain regions of normal and hypertensive rats in the presence and absence of gamma-2-MSH and/or ACTH[4-10]; (2) protein phosphorylation immunoprecipitation and Western immunoblot assays to determine whether phorbol esters, in the presence and absence of gamma-2-MSH and ACTH[4-10] interfere with the activation of PKC-delta, -epsilon, and-gamma synaptosomes from the forebrain, midbrain, and hindbrain regions of normal and hypertensive rats; and (3) calcium imaging studies to determine whether phorbol esters increase or decrease intracellular Ca2+ levels in synaptosomes isolated from the forebrain, midbrain, and hindbrain regions of normal and hypertensive rats in the presence and absence of gamma-2-MSH and ACTH[4-10].

描述（改编自申请）：磷酸肌醇信号传导 据报道，系统和钙在调节中发挥作用 血压。在初步研究中，已建立 免疫印迹和免疫组织化学研究表明 PKC 同工酶 α、 beta、gamma、delta、epsilon 和 zeta 在突触体中表达 从正常和高血压大鼠的大脑区域分离。它还具有 已确定 gamma-2-MSH 改变 PKC 同工酶的表达 以及高血压大鼠的 PKC 活性。据报道 来自细胞外空间的 ACTH 样肽影响 B-50 (GAP-43) 磷蛋白通过与 神经末梢的突触前膜。而且脑子也很发达 PKC和Ca2+对高血压敏感，导致多种疾病 脑血管和神经元的变化。综合起来就是 假设在高血压中，肽之间存在联系 （即 γ-2-MSH、ACTH[4-10] 和 ACTH] PKC 介导的磷酸化 特定 PKC 同工酶的变化，导致 PKC 介导的变化 特定蛋白质的磷酸化以及 钙含量。 为了了解中枢神经系统的作用和 研究人员发现高血压中的磷酸肌醇信号通路 计划使用：（1）蛋白质磷酸化、免疫沉淀和 蛋白质免疫印迹分析可鉴定特定蛋白质 由于 PKC-delta、-epsilon 激活而磷酸化， 正常人前脑、中脑和后脑区域的 和-γ 以及存在或不存在 γ-2-MSH 和/或的高血压大鼠 促肾上腺皮质激素[4-10]； (2)蛋白磷酸化免疫沉淀及Western印迹 免疫印迹测定以确定佛波酯是否存在 并且缺少 gamma-2-MSH 和 ACTH[4-10] 会干扰激活 来自前脑的 PKC-delta、-epsilon 和-gamma 突触体， 正常和高血压大鼠的中脑和后脑区域；和（3） 钙成像研究以确定佛波酯是否增加或 降低分离自突触体的细胞内 Ca2+ 水平 正常和高血压的前脑、中脑和后脑区域 存在和不存在 γ-2-MSH 和 ACTH 的大鼠[4-10]。