ULTRASTRUCTURE OF THE MYOCARDIAL SARCOLEMMA

心肌肌膜的超微结构

• 批准号: 3340076
• 负责人: JOY S FRANK
• 金额: $ 17.43万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1982
• 资助国家: 美国
• 起止时间: 1982-05-01 至 1995-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3340076
• 关键词: anesthetics; calcium transporting ATPase; cell membrane; digitalis; extracellular matrix; freeze etching; glycolipids; glycoproteins; heart pharmacology; histochemistry /cytochemistry; immunoelectron microscopy; laboratory rabbit; laboratory rat; membrane activity; membrane permeability; myocardial ischemia /hypoxia; myocardium; phospholipase D; sarcolemma; verapamil

This is a study of the myocardial sarcolemma and the extracellular matrix. This project attempts to gain an understanding of the function of the myocardial sarcolemma from an ultrastructural analysis of the components of the cell surface (glycocalyx) in the bilayer, their relationship to each other, to the extracellular matrix and to membrane transport. The approach is to study the macromolecular architecture of the bilayer and glycocalyx/extracellular matrix with state-of-the-are freeze-fracture and immunoelectron microscopy. The overall aims are to determine (1) if any of the intramembrane particles as revealed in freeze-fracture and freeze-etch membrane are related to ion transport and (2) which specific macromolecules within the glycocalyx have an important structural and functional relationship to the bilayer and to components of he extracellular matrix. The methodology is to (1) examine the filament of the glycocalyx and extracellular matrix to determine in untreated tissue their configuration and identity. This will be performed in normal hearts, during development and after brief periods of ischemia, (2) to determine the distribution and localization in the bilayer of two major transport proteins, i.e. Na,K- ATPase and Na-Ca exchanger. A combination of the following techniques will be used on isolated adult myocytes, cultured myocytes and heart tissue: ultra-rapid freezing, freeze-drying, freeze-substitution, low temperature embedding, freeze-fracture and etch, antibody and immunogold labeling and label fracture. The proposed research has a two-fold significance, (1) by identifying structural components of the glycocalyx/extracellular matrix and intramembrane particles int he bilayer, with specific membrane function it addresses a question fundamental to the biology of all cells and (2) it utilizes stat-of-the-art ultrastructure to focus on structure-function relationships (both physiological and pathological) of the sarcolemma and extracellular matrix, where in the heart there is little information.

这是对心肌肌膜和细胞外基质的研究。 该项目试图了解 心肌肌膜成分的超微结构分析 双层中的细胞表面（糖萼），它们与每个的关系 其他，到细胞外基质和膜运输。 方法 是研究双层的大分子结构 具有现状冷冻断裂的糖萼/细胞外基质和 免疫电子显微镜。 总体目标是确定 (1) 是否存在以下任一情况： 冷冻断裂和冷冻蚀刻中显示的膜内颗粒 膜与离子传输有关，以及 (2) 哪些特定的大分子 糖萼内具有重要的结构和功能 与双层和细胞外基质成分的关系。 该方法是（1）检查糖萼的细丝和 细胞外基质以确定未处理组织中的结构 和身份。 这将在正常心脏的发育过程中进行 并在短暂的缺血后，（2）确定分布和 定位于两种主要转运蛋白的双层，即 Na,K- ATP 酶和 Na-Ca 交换器。 以下技术的组合将 用于分离的成体肌细胞、培养的肌细胞和心脏组织： 超快速冷冻、冷冻干燥、冷冻替代、低温 包埋、冷冻断裂和蚀刻、抗体和免疫金标记以及 标签断裂。 拟议的研究具有双重意义，（1）通过确定 糖萼/细胞外基质的结构成分和 膜内颗粒位于双层中，具有特定的膜功能 解决了所有细胞生物学的基本问题，并且 (2) 利用最先进的超微结构来关注结构功能 肌膜的关系（生理和病理）和 细胞外基质，心脏中的信息很少。