ULTRASTRUCTURE AND CHARGE PERMSELECTIVITY OF ENDOTHELIUM

内皮的超微结构和电荷选择性

• 批准号: 3343822
• 负责人: GIUSEPPE G PIETRA
• 金额: $ 15.76万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 1984
• 资助国家: 美国
• 起止时间: 1984-08-01 至 1992-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3343822
• 关键词: basement membrane; blood; electron microscopy; electron probe spectrometry; histochemistry /cytochemistry; lung; lymph; macromolecule; microcirculation; mucopolysaccharides; peptidases; perfusion; pulmonary circulation; pulmonary edema; pulmonary surfactants; radiotracer; tissue /cell culture; vascular endothelium permeability

This proposal aims to define the contribution of the pulmonary microvascular endothelium to size and charge permselectivity in the lung. The hypothesis to be tested is that anionic glycoproteins in glycosaminoglycans on the surface of the plulmonary microvascular endothelium are organized in discrete domains which may influence the routes taken by macromolecules across the capillary wall. Pathways for neutral and charged macromolecules across the capillary walls will be identified qualitatively by electron microscopy using electron-dense permeability probes of different molecular size and net electrical charge. The experiments will be done in the isolated perfused rat lung preparation to avoid systemic effects resulting from interaction between charged probes and blood constituents. Quantitative assessment of transcapillary movement of permeability probes will be done by morphometric measurements at different time intervals and/or by measurements of lung radioactivity in lungs perfused with radioactive permeability probes. Selective enzymatic removal of glycoproteins or glycosaminoglycans on the endothelium will provide insights in the biochemical nature of the constituents of the charged barrier. Using lectins bound to colloidal gold the distribution of carbohydrate units of different glycoproteins on the microvascular endothelium and adjacent basement membrane will be examined in thin sections of lungs. The overall goal of the project is to identify new structural determinants of lung permeability and gain information for future studies on mechanism of endothelial damage in differnt types of lung injury.

该建议旨在定义肺的贡献 微血管内皮到肺中的尺寸和电荷宽度率。 要测试的假设是 毛糖胺聚糖表面的糖胺聚糖 内皮是在离散域中组织的，这可能会影响 大分子在整个毛细管壁上采用的路线。 途径 毛细管壁上的中性和带电的大分子将是 通过电子密集的电子显微镜定性鉴定 分子大小和净电荷的渗透率概率。 实验将在孤立的灌注大鼠肺制备中进行 为了避免由带电探针之间的相互作用产生的全身效应 和血液成分。 跨毛皮运动的定量评估 渗透率概率将通过形态计量测量来完成 不同的时间间隔和/或通过测量肺放射性 肺有放射性通透性探针。 选择性酶促 去除内皮上的糖蛋白或糖胺聚糖将 提供有关成分的生化性质的见解 充电障碍。 使用与胶体黄金结合的凝集素的分布 微血管上不同糖蛋白的碳水化合物单元 内皮和邻近的地下膜将在薄中检查 肺部。 该项目的总体目标是确定新的 肺通透性的结构决定因素并获得信息 未来关于内皮损害机理的肺部损害机制 受伤。