CHLORIDE AND OSMOLYTES AND VOLUME REGULATION

描述：细胞体积调节对整体细胞生长至关重要。 内稳态，并在各种病理情况下发挥作用， 昏迷水肿中风和糖尿病 本提案采用以下组合 电生理、荧光和共聚焦显微镜和分子生物学 生物技术来解决阴离子，渗透剂的生理作用 和细胞骨架的调节。 重点是一个 体积调节阴离子电导（VRAC），这是一个无处不在的成员， 机械敏感离子通道家族。 这种电导在一个 广泛的细胞类型，并已涉及从细胞的恢复 肿胀在许多细胞，包括淋巴细胞，模型系统中使用的 这些研究。 这一建议涉及几个假设，旨在了解 调节这种体积调节阴离子电导和贡献 通过同步测量， 电流幅度和细胞肿胀的变化。 具体解决的是假设，体积调节阴离子电流 活性受细胞内的组成和浓度调节， 渗透剂 电流和体积测量将结合分子 生物技术来解决这一假设， 电流也由小的细胞溶质肌动蛋白结合蛋白调节， 称为ICln。 肌动蛋白细胞骨架的变化为细胞提供 结构也涉及调节细胞体积， 被认为是激活阴离子电流的机械换能器。 这 假设将解决两种蛋白质，BotC 2和凝溶胶蛋白，以改变 F-和G-肌动蛋白之间的平衡，并确定对 电流激活和细胞肿胀使用电生理学，分子 和荧光技术。 成功完成此提案将 更深入地了解 细胞中的机械力转导及其在压力下的体积调节中的作用 正常和病理状态。

DESCRIPTION: Cell volume regulation is critical to overall cellular homeostasis and plays a role in a variety of pathological situations such as coma, edema, stroke and diabetes. This proposal uses a combination of electrophysiological, fluorescent and confocal microscopy and molecular biology techniques to address the physiological role of anions, osmolytes and the cytoskeleton in volume regulation. The focus is on a volume-regulated anion conductance (VRAC), which is a member of a ubiquitous family of mechanosensitive ion channels. This conductance is found in a wide variety of cell types and has been implicated in the recovery from cell swelling in many cells, including lymphocytes, the model system used in these studies. This proposal addresses several hypotheses directed toward understanding the regulation of this volume-regulated anion conductance and the contribution of this conductance to cellular volume regulation by concurrent measurements of current amplitude and cell swelling in a hyposmotic environment. Specifically addressed is the hypothesis that volume regulated anion current activity is regulated by the intracellular composition and concentration of osmolytes. Current and volume measurements will be combined with molecular biological techniques to address the hypothesis that the activity of this current is also regulated by a small, cytosolic actin binding protein, termed ICln. Changes in the actin cytoskeleton which provide cells with structure have also been implicated in regulating cell volume and have been suggested as the mechanotransducer which activated the anion current. This hypothesis will be addressed with two proteins, BotC2 and gelsolin to alter the equilibrium between F- and G- actin and determining the effect on current activation and cell swelling using electrophysiological, molecular and fluorescent techniques. Successful completion of this proposal will provide a greater understanding of the underlying mechanisms of mechanotransduction in cells and their role in volume regulation under normal and pathological conditions.