GENERATION AND REGULATION OF FORCE IN SMOOTH MUSCLE

平滑肌力的产生和调节

• 批准号: 3334789
• 负责人: FREDRIC S FAY
• 金额: $ 29.97万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-09-30 至 1990-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3334789
• 关键词: Anura; calcium; cyclic AMP; fresh water environment; guinea pigs; high energy compound; ion transport; mechanical pressure; membrane potentials; muscle cells; muscle contraction; muscle tension; neuromuscular transmission; neurotransmitters; newborn animals; oxygen tension; single cell analysis; smooth muscle

Studies will be performed to determine the manner in which contractile proteins are organized into a contractile apparatus in smooth muscle, and how changes in this order might account for active cell shortening as well as the length dependence of active force development. Studies will also be performed to characterize the mechanical events underlying force development, and to analyze the role played by Ca+2 in regulating these processes. These studies will all be performed on single isolated smooth muscle cells, thereby avoiding the complexities of intact multicellular preparations and affording a direct view of cellular properties. Single smooth muscle cells will be isolated by enzymatic disaggregation of the stomach of Bufor marinus. The distribution of contractile proteins will be analyzed using flourescently labelled protein specific probes in both fixed and living cells. Fluorescent tags will be attached to either antibodies (both mono- and polyclonal) or purified native contractile protein; the former will be used to localize contractile protein in fixed cells and either one of these may be used in living cells into which they will be introduced by microinjection. The distribution of fluorescence will be assessed utilizing a low light level TV whose output is subject to digital image processing to obtain a computerized reconstruction of the distribution of contractile protein within a single isolated smooth muscle cell. Contractile state of these cells will be monitored by isometric measurement of force by tying a single cell to a force transducer. Ca+2 will be measured using a new class of highly fluorescent Ca+2 indicators which are trapped inside the smooth muscle cell by the action of cellular esterases. The proposed studies employing tools of biophysics, biochemistry, cell biology and computer science are part of long term effort to determine the mechanisms underlying the generation and regulation of force in smooth muscle. These basic studies of normal function of smooth muscle should provide much needed insight into derangements of smooth inside function in diseases such as hypertension, asthma, and spastic disorders of the G. I. system.

将进行研究以确定收缩的方式 蛋白质被组织成平滑肌的收缩仪 该顺序的变化如何也解释了主动电池缩短 作为主动力发展的长度依赖性。 研究也将是 进行表征的机械事件的表征 开发，并分析CA+2在调节这些方面的作用 过程。 这些研究将全部在单个孤立的平滑上进行 肌肉细胞，从而避免了完整的多细胞的复杂性 制备并提供细胞特性的直接视图。 单身的 平滑肌细胞将通过酶促分解来分离 Bufor Marinus的胃。 收缩蛋白的分布将是 在两个固定的两个固定的蛋白质特异性探针中使用粉状标记的蛋白质特异性探针进行了分析 和活细胞。 荧光标签将连接到两种抗体 （单单和多克隆）或纯化的天然收缩蛋白；这 前者将用于将收缩蛋白定位在固定细胞中，并且 其中一个都可以用于将其进入的活细胞中 由微注射引入。 荧光的分布将是 利用低光电视的评估，其输出受数字的影响 图像处理以获得计算机化的重建 单个孤立平滑肌内收缩蛋白的分布 细胞。 这些单元的收缩状态将通过等距监测 通过将单个细胞与力传感器绑定到力来测量力。 CA+2 将使用新的高荧光CA+2指标测量 通过细胞的作用将其捕获在平滑肌细胞内 酯酶。 拟议的研究采用生物物理学工具， 生物化学，细胞生物学和计算机科学是长期的一部分 努力确定发电和调节的基础机制 平滑肌的力。 这些对正常功能的基础研究 平滑肌应提供急需的洞察力 在高血压，哮喘和 G. I.系统的痉挛疾病。