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

描述（改编自应用）：磷酸肌醇信号转导 系统和钙已被报道在调节中发挥作用 血压。在初步研究中， 免疫印迹和免疫组织化学研究PKC同工酶α， β、γ、δ、δ和ζ在突触体中表达 分离自正常和高血压大鼠的脑区域。它还 已经确定γ-2-MSH改变PKC同工酶的表达 以及高血压大鼠PKC活性。据报 促肾上腺皮质激素样肽来源于细胞外空间， B-50（GAP-43）磷蛋白通过与 神经末梢的突触前膜。而且，大脑是里克在 PKC与Ca ~（2+）的关系密切，对高血压敏感，高血压可引起多种 血管和神经元大脑变化。综合来看， 假设在高血压中， (i.e.,γ-2-MSH、ACTH[4-10]和ACTH] PKC介导的磷酸化 特异性PKC同工酶，这导致PKC介导的 特定蛋白质的磷酸化以及 钙的水平。 为了了解中枢神经系统的作用， 高血压中的磷酸肌醇信号通路， 计划用途：（1）蛋白磷酸化，免疫沉淀， 蛋白质免疫印迹分析，以确定特定的蛋白质， 由于PKC-δ，-β， 正常人前脑、中脑和后脑区域中的 和高血压大鼠在存在和不存在γ-2-MSH和/或 ACTH[4-10];（2）蛋白磷酸化免疫沉淀和Western 免疫印迹分析以确定佛波醇酯在存在下 而γ-2-MSH和ACTH的缺乏[4-10]干扰激活 PKC-δ、PKC-γ和PKC-γ突触体的数量， 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 降低分离的突触体细胞内Ca 2+水平， 正常和高血压的前脑、中脑和后脑区域 大鼠在存在和不存在γ-2-MSH和ACTH的情况下[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].