GENERATION AND REGULATION OF FORCE IN SMOOTH MUSCLE

平滑肌力的产生和调节

• 批准号: 3569244
• 负责人: FREDRIC S FAY
• 金额: $ 43.53万
• 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 2000-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3569244
• 关键词: alternatives to animals in research; asthma; bioenergetics; calcium; fresh water environment; high energy compound; hypertension; ion transport; mechanical pressure; membrane potentials; muscle cells; muscle contraction; muscle tension; neuromuscular transmission; neurotransmitters; protein kinase C; single cell analysis; smooth muscle; toad

(Adapted from the Investigator's abstract) Studies will be performed to determine how Ca2+ is regulated in smooth muscle and how in turn Ca2+ regulates contraction of these cells. To this end studies will be performed to characterize the mechanisms responsible for the storage and release of Ca2+ from internal storage compartments. The spatial organization of these compartments and their functional and structural interconnection will be determined. Sites of Ca2+ entry into cells, as well as mechanisms linking transmembrane Ca2+ entry with release of Ca2+ from internal stores will be determined. Studies into the mechanisms responsible for removing Ca2+ from the cell, and for modulating the activity of these processes will be carried out. The role of protein kinase C as a feedback regulator of Ca2+ signalling mechanisms will also be determined. These studies will be performed on single isolated smooth muscle cells, thereby avoiding the complexities of intact multicellular preparations and affording a direct view into these cellular/subcellular issues. Single smooth muscle cells will be isolated by enzymatic disaggregation of the stomach muscularis of the toad, Bufo Marinus. This system is a well=characterized model system for studying smooth muscle physiology at the cellular level. The properties of Ca2+ transport and release by internal compartments will be investigated using a saponin skinned preparation measuring ion fluxes by techniques involving isotopes or fluorescent indicators. The 3D molecular cytoarchitecture of these compartments, will be determined by fluorescent antibodies in conjunction with the digital imaging microscope. Sudden changes in the level of key molecules suspected to be involved in Ca2+ regulation in smooth muscle will be accomplished by their photolytic generation from caged precursors. The role of protein kinases will be tested by injection of peptide modulators of specific kinases. The Ca2+ sensitivity of activation of smooth muscle contraction will be assessed by simultaneous measurements of Ca2+ and force in a single smooth muscle cell. The proposed studies employing tools of biophysics, molecular biology, cell biology and computer science are part of a long-term effort to determine the mechanism 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 muscle function in diseases such as hypertension, asthma, and spastic disorders of the G.I. System.

（根据调查员的摘要改编）将进行研究 确定如何在平滑肌中调节Ca2+以及如何依次CA2+ 调节这些细胞的收缩。 为此，最终研究将是 执行以表征负责存储的机制和 从内部存储室释放Ca2+。 空间 这些隔室的组织及其功能和结构 将确定互连。 Ca2+进入细胞的位置，AS 以及将跨膜CA2+进入CA2+的机制连接起来 将从内部商店确定。 研究机制 负责从单元中删除Ca2+，并调制 这些过程的活动将进行。 蛋白质的作用 激酶C作为CA2+信号传导机制的反馈调节剂也将 可以确定。 这些研究将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 制备并直接看到这些细胞/亚细胞 问题。 单个平滑肌细胞将通过酶促分离 蟾蜍bufo marinus的胃肌肉的分解。 这 系统是一个井=用于研究平滑肌的特征模型系统 细胞水平的生理学。 Ca2+运输的特性和 通过皂苷将通过内部隔室释放 通过涉及同位素的技术测量离子通量的皮肤制备 或荧光指标。 这些的3D分子细胞结构结构 隔室将通过荧光抗体结合确定 使用数字成像显微镜。 钥匙水平的突然变化 涉嫌参与平滑肌中Ca2+调节的分子 将通过笼中的前体的光解生成来实现。 蛋白激酶的作用将通过注射肽进行测试 特定激酶的调节剂。 CA2+激活的灵敏度 平滑肌收缩将通过同时测量来评估 Ca2+和单个平滑肌细胞中的力。 提出的研究 采用生物物理学，分子生物学，细胞生物学和 计算机科学是确定机制的长期努力的一部分 基础平滑肌中力的产生和调节。 这些 平滑肌正常功能的基础研究应提供很多 需要深入了解疾病中平滑肌功能的危险 例如高血压，哮喘和G.I.的痉挛性疾病系统。

项目成果

Role of Na(+)-Ca2+ exchanger in beta-adrenergic relaxation of single smooth muscle cells.

Na( )-Ca2 交换剂在单个平滑肌细胞 β-肾上腺素能松弛中的作用。

• DOI: 10.1111/j.1749-6632.1991.tb17347.x
• 发表时间: 1991
• 期刊: Annals of the New York Academy of Sciences
• 影响因子: 5.2
• 作者: Moore,ED;Fogarty,KE;Fay,FS
• 通讯作者: Fay,FS

Calcium transients and resting levels in isolated smooth muscle cells as monitored with quin 2.

• DOI: 10.1152/ajpcell.1986.250.5.c779
• 发表时间: 1986-05
• 期刊: The American journal of physiology
• 作者: D. Williams;F. Fay

Contractile mechanism of single isolated smooth muscle cells.

单个分离的平滑肌细胞的收缩机制。

• 发表时间: 1982
• 期刊: Society of General Physiologists series
• 作者: Fay,FS;Fogarty,K;Fujiwara,K;Tuft,R
• 通讯作者: Tuft,R

Isoproterenol stimulates rapid extrusion of sodium from isolated smooth muscle cells.

异丙肾上腺素刺激钠从分离的平滑肌细胞中快速排出。

• DOI: 10.1073/pnas.90.17.8058
• 发表时间: 1993
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Moore,ED;Fay,FS
• 通讯作者: Fay,FS

Rectifying properties of the membrane of single freshly isolated smooth muscle cells.

单个新鲜分离的平滑肌细胞膜的整流特性。

• DOI: 10.1152/ajpcell.1980.239.5.c175
• 发表时间: 1980
• 期刊: The American journal of physiology
• 作者: Singer,JJ;WalshJr,JV
• 通讯作者: WalshJr,JV