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MEMBRANE PERTURBATION DURING INFLAMMATION

炎症期间的膜扰动

基本信息

批准号：
3426269
负责人：
HERBERT E WHITELEY
金额：
$ 1.96万
依托单位：
UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
依托单位国家：
美国
项目类别：
财政年份：
1985
资助国家：
美国
起止时间：
1985-09-30 至 1987-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3426269
关键词：
biological models carbohydrate structure cell adhesion cell migration cell type cellular pathology corneal endothelium freeze etching inflammation intercellular connection leukocytes membrane structure model design /development scanning electron microscopy

项目摘要

This proposal is in response to the small grants program for recently trained, less experienced basic scientists. The specific aims and long-term objectives are to develop the bovine corneal cup as a method for studying changes in membrane during the inflammatory process. This method will allow dissection of the inflammatory cell-corneal endothelial cell interaction from a morphologic and functional viewpoint. Many ramifications of the inflammatory process within the eye can eventually be explored by use of specific cell types, stimulated cells (i.e., activated neutrophils, lymphocytes, macrophages), specific humoral or chemical mediators of inflammation, viral or bacterial disease states, and blocking receptors such as lectins. Several specific questions will be addressed in this initial study: 1) What are the changes in corneal endothelial cell surface morphology and carbohydrate structure during inflammatory cell adhesion and transmigration?; 2) Are there changes in the morphology or function of endothelial intercellular junctions?; and 3) Are these changes dependent on a specific inflammatory cell type? In order to begin answering these questions the bovine corneal cup model will be established with several techniques being applied to the model. Specific peripheral blood leukocytes will be isolated. Changes in endothelial cell surface morphology will be determined by the use of scanning electron microscopy and the freeze-fracture technique with quantitative data accumulated by determination of intramembranous particle size and density distribution. Changes in cell surface carbohydrate structure will be determined with a battery of lectins complexed with hemocyanin which then can be quantitated by use of scanning electron microscopy. Changes in intercellular junctions will be identified by quantitative assessment of junction size and distribution by use of the freeze-fracture technique. Establishment of the corneal cup model will allow a more controlled and detailed examination of inflammatory cell-endothelial cell interaction.

这项建议是为了响应最近的小额赠款计划，训练有素经验不足的基础科学家具体目标和长期目标是开发牛角膜杯作为研究炎症过程中细胞膜的变化。该方法将允许解剖炎性细胞角膜内皮细胞从形态学和功能学的角度来看，许多眼睛内炎症过程的分支最终会通过使用特定的细胞类型，刺激的细胞（即，激活嗜中性粒细胞、淋巴细胞、巨噬细胞），特异性体液或化学炎症介质、病毒或细菌疾病状态和阻断受体如凝集素。在本初步研究中将讨论几个具体问题：1）角膜内皮细胞表面形态和炎症细胞粘附过程中的碳水化合物结构，轮回？2)是否有形态或功能的变化，内皮细胞间连接;（3）这些变化是否取决于一种特定的炎症细胞为了开始回答这些问题，将通过应用于模型的几种技术来建立。将分离特定的外周血白细胞。变化内皮细胞表面形态将通过使用扫描电子显微镜和冷冻断裂技术通过测定膜内颗粒积累的定量数据尺寸和密度分布。细胞表面糖的变化结构将用一组凝集素与然后可以通过使用扫描电子定量血蓝蛋白显微镜细胞间连接的变化将通过定量评估结的大小和分布的使用冷冻断裂技术角膜杯模型的建立将允许更可控的和炎症细胞-内皮细胞相互作用的详细检查。