FUNCTION OF GUT PEPTIDES ISOLATED GUT MUSCLE CELLS

肠肽分离肠肌细胞的功能

• 批准号: 3228711
• 负责人: GABRIEL M. MAKHLOUF
• 金额: $ 10.23万
• 依托单位: VIRGINIA COMMONWEALTH UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-04-01 至 1989-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3228711
• 关键词: acetylcholine; adenosine triphosphate; calcium metabolism; cell membrane; cholecystokinin; combination chemotherapy; cyclic AMP; cyclic GMP; cyclic nucleoside monophosphate; cytoplasm; gallbladder; gastrins; gastrointestinal hormones; human tissue; intestines; ion transport; ionophores; kinins; lanthanum; membrane permeability; muscle cells; muscle contraction; muscle proteins; muscle relaxation; naloxone; neurotransmitters; opiate alkaloid; peptide hormone; peptides; phenothiazines; phosphodiesterases; phosphorylation; physical chemical interaction; radiotracer; smooth muscle; stoichiometry; stomach; vasoactive intestinal peptide; verapamil

The objective of this proposal is to characterize the properties of smooth muscle cells isolated from the mammalian gut. We have developed techniques for isolation of muscle cells from the stomach (guinea pig, dog and man) and gallbladder (dog and man) and have devised an image-splitting micrometric technique for the direct measurement of contraction and relaxation. We have also applied techniques for the indirect measurement of muscle cell response in terms of cellular intermediates: intracellular levels of cyclic nucleotides; 45Ca ions efflux and the phosphorylated fraction of myosin light chain (using IEF/SDS electrophoresis). The specific aims of the proposal are: 1) to identify and characterize smooth muscle receptors for gut peptides; these are known to be present in nerve fibers of the submucous and myenteric plexuses of the gut and are thought to play a physiological role as neurotransmitters or neuromodulators in the control of smooth muscle activity; the most likely candidates are VIP, the opioid peptides (met- and leu-enkephalin), substance P and bombesin. 2) To define the contractile response to peptides by measurement of 45Ca ions efflux and the phosphorylated fraction of myosin light chain, and 3) To define the role of cyclic AMP in smooth muscle relaxation induced by VIP and its homologues and test the possibility that ATP (the putative 'purinergic' relaxant neurotransmitter) and VIP interact synergistically to modulate neurally-induced relaxation.

该提议的目的是表征平滑的特性 从哺乳动物肠道中分离出来的肌肉细胞。 我们已经开发了针对的技术 从胃中隔离肌肉细胞（豚鼠，狗和人）， 胆囊（狗和男人），并设计了一个图像切开的微米 直接测量收缩和放松的技术。 我们有 还应用技术来间接测量肌肉细胞反应 细胞中间体的术语：循环核苷酸的细胞内水平； 45CA离子外排和肌球蛋白轻链的磷酸化部分（使用 IEF/SDS电泳）。 该提案的具体目的是：1）确定 并表征肠肽的平滑肌受体；这些是 存在于肠道粘膜和肌丛的神经纤维中， 被认为是神经递质或神经调节剂的生理作用 控制平滑肌活动；最有可能的候选人是VIP， 阿片类肽（Met-和Leu-enkephalin），物质P和孟买。 2）通过测量45CA离子来定义对肽的收缩反应 外排和肌球蛋白轻链的磷酸化部分，3）定义 循环AMP在VIP及其引起的平滑肌松弛中的作用 同源物并测试ATP（推定的“嘌呤能”放松剂的可能性 神经递质）和VIP协同相互作用以调节神经诱导的 松弛。