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

说明(改编自应用)：肌醇磷脂信号转导 据报道，系统和钙在调节中发挥了作用。 血压的变化。在初步研究中，它已经由 免疫印迹和免疫组织化学研究PKC同工酶α， β、伽马、德尔塔、epsilon和zeta在突触体中表达 从正常大鼠和高血压大鼠的脑区分离。它还 已证实γ-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)磷酸蛋白通过与 神经末梢的突触前膜。此外，大脑也被激活了 PKC和Ca~(2+)对引起多种疾病的高血压敏感 脑血管和神经元的变化。综上所述，它是 假设在高血压中，多肽之间存在联系 (即，γ-2-MSH、ACTH[4-10]和ACTH]PKC介导的磷酸化 特定的PKC同工酶，从而导致PKC介导的变化 特定蛋白质的磷酸化以及基因的改变 钙的水平。 为了了解中枢神经系统的作用和 高血压中的磷酸肌醇信号通路研究进展 计划使用：(1)蛋白质磷酸化、免疫沉淀和 免疫印迹分析鉴定特定的蛋白质 作为PKC-Delta，-epsilon活化的结果而被磷酸化， 正常人前脑、中脑和后脑区域的-GAMA 和高血压大鼠在存在和不存在γ-2-MSH和/或 ACTH[4-10]；(2)蛋白质磷酸化免疫沉淀与Western 免疫印迹法检测佛波酯是否存在 而缺乏γ-2-MSH和ACTH[4-10]则干扰激活 来自前脑的PKC-Delta、-epsilon和-Gamma突触体， 正常大鼠和高血压大鼠的中脑和后脑区域； 钙成像研究，以确定佛波酯是否增加或 降低分离的突触体内钙离子水平 正常和高血压患者的前脑、中脑和后脑区域 在存在和不存在γ-2-MSH和ACTH的情况下的大鼠[4-10]。

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].