HORMONAL AND IONIC CONTROL OF METABOLISM

拟议研究的主要目的是更全面地界定 钙离子调控细胞功能的机制 如激素和离子调节等过程 肾上腺肾小球细胞分泌的醛固酮。细胞 培养的3T3细胞的增殖和白细胞功能。 具体目标包括阐明时间模式 蛋白质磷酸化、肌醇脂代谢、钙离子 血管紧张素II过程中的代谢和蛋白激酶C功能 和蛙皮素分别在肾小球和肾小球中介导的反应 3T3细胞。II型钙调素依赖性蛋白激酶在细胞周期调控中的作用 细胞反应的初始阶段；免疫细胞化学 C-激酶的定位及M-激酶在脑出血中的作用 持续的细胞反应；信息传递的机制 通过C-激酶从细胞表面传递到细胞内部 钙离子信使系统在持续期的分支 细胞反应特别关注Na+/H+的变化 交换和可能的蛋白激酶级联；分子 细胞C-激酶介导期的记忆基础 回应；以及对事件时间模式的重新审视 ACTH介导的醛固酮在cAMP信使系统中的作用 肾上腺肾小球细胞分泌的系统 来探讨这些问题。方法将包括对钙离子的研究 两种方法测定细胞内游离钙离子的通量 Fura 2和Aequorin作为指标；研究 肌醇脂类和肌醇磷酸盐的化学和放射性- 同位素方法.磷蛋白的二维凝胶电泳法 从细胞提取液中获得；光和EM C-激酶的免疫细胞化学定位。下面的问题 调查具有广泛的生物学意义，并具有重要意义。 对于广泛的生物现象。更好地理解 钙信使系统功能的研究可能会改变我们对 细胞生长和增殖、多肽和类固醇激素 分泌，和平滑肌肉收缩。

The major aims of the proposed research is to more fully define the mechanisms by which CA2+ regulates cell function during such processes as the hormonal and ionic regulations of aldosterone secretion, from adrenal glomerulosa cells. Cell proliferation of cultured 3T3 cells, and leukocytes function. Specific aims include an elucidation of the temporal patterns of protein phosphorylation, inositol lipid metabolism, Ca2+ metabolism, and protein kinase C function during angiotensin II and bombesin-mediated response in, respectively, glomerulosa and 3T3 cells. The role of type II CaM-dependent protein kinase in the initial phase of cellular response; the immunocytochemical localization of the C-kinase and the possible role of M-kinase in sustained cellular response; the mechanism by which information is conveyed from cell surface to cell interior via the C-kinase branch of the Ca2+ messenger system during the sustained phase of cellular response focusing particularly on changes in Na+/H+ exchange and possible protein kinase cascades; the molecular basis of memory in the C-kinase-mediated phase of cellular response; and a reexamination of the temporal pattern of events in the cAMP messenger system during ACTH-mediate aldosterone secretion from adrenal glomerulosa cells as the system in which to explore these issues. Methods will include studies of Ca2+ fluxes and measurements of intracellular free Ca2+ using both fura 2 and aequorin as indicators; studies of the turnover of inositol lipids and inositols phosphates by both chemical and radio- isotope methods; 2-D gel electrophoresis of the phosphoproteins obtained from extracts of cells; and both light and EM immunocytochemical localization of C-kinase. The problem under investigation is of broad biological interest, and has implications for a wide range of biological phenomena. A better understanding of Ca2+ messenger system function is likely to alter our views of cell growth and proliferation, peptide and steroid hormone secretion, and smooth muscle contraction.