CELLULAR BIOLOGY OF RENAL FUNCTION AND DISEASE

该计划项目的目标是研究蜂窝 肾上皮细胞膜功能和功能障碍的生物学研究 和肾小球细胞。战略将是使用融合、 超微结构免疫化学的先进技术，冷冻- 断裂、分子生物学、生物物理和生化 探索不同细胞类型中相互关联的问题。 具体地说，膜回收和微丝的影响 对加压素诱导的水流量的调节将被研究。 我们将研究笼状蛋白包被的小凹和囊泡在 膜水通道的循环利用，并尝试 水通道蛋白的分离、纯化和鉴定(S)。这个 微丝在跨上皮水流动中的相互作用 将在完整的细胞中进行研究，并在体外与 将评估纯化的肌动蛋白和肌动蛋白相关的系统 蛋白质相互作用及其凝胶-溶胶化能力 在渗透梯度存在和不存在的情况下的转化。 一种已定义的蛋白质H+ATPase的膜循环将是 在质子分泌细胞中进行检查，利用 抗H+ATPase的单抗。两国之间的关系 H+-ATPase分子的其他特征膜特性， 包括独特的非分子筛蛋白包衣小泡的转运，将是 检查过了。这些研究将得到以下研究的补充： 肾细胞功能障碍。细胞和分子 海曼病发病机制中的抗原生物学 将对肾炎进行检查，以确定脱落与 引起肾炎的抗原gp330的内吞作用是在 肾小球上皮细胞。肾小球系膜细胞功能 将通过确定炎症介质是否 激活系膜细胞中的钙反应，进而 促进收缩、增殖和前列腺素的产生 这可能与糖尿病肾病中的类似现象有关。 通过探索一些一般的细胞生物学过程 与膜的动态调节密切相关 功能，我们希望相互作用可以在分子上定义 水平。这项研究计划涉及的所有项目都分享 这个共同的主题。很可能是细胞中的一些问题 生物学是细胞功能正常和不正常的关键 回答，加深了我们对肾功能和 疾病。

The aim of this Program Project is to investigate the cellular biology of membrane function and dysfunction in renal epithelial and glomerular cells. The strategy will be to use convergent, advanced techniques in ultrastructural immunochemistry, freeze- fracture, molecular biology, biophysics and biochemistry to explore interrelated problems in different cell types. Specifically, membrane recycling and the effect of microfilament modulation on vasopressin-induced water flow will be examined. We will investigate the role of clathrin coated pits and vesicles in the recycling of membrane water channels, and attempt to isolate, purify and characterize the water channel protein(s). The interactive role of microfilaments in transepithelial water flow will be studied in intact cells and correlations made with in vitro systems which will evaluate purified actin and actin-associated protein interactions and their ability to undergo gel-sol transformation in the presence and absence of osmotic gradients. Membrane recycling of a defined protein, the H+ATPase, will be examined in proton secreting cells, taking advantage of a monoclonal antibody against H+ATPase. The relationship of H+ATPase molecules to other characteristic membrane features, including transport by unique nonclathrin coated vesicles, will be examined. These studies will be complemented by studies on cellular dysfunction of renal cells. The cellular and molecular biology of antigens involved in the pathogenesis of Heymann nephritis will be examined to determined how shedding versus endocytosis of a nephritogenic antigen, gp330, is achieved in glomerular epithelial cells. Glomerular mesangial cell function will be probed by determining if mediators of inflammation activate a calcium response in the mesangial cell, which in turn promotes contraction, proliferation, and prostanoid production that can be related to similar phenomona in diabetic nephropathy. By probing a number of general cell biological processes intimately involved with the dynamic regulation of membrane function, we hope to interactions can be defined at the molecular level. All of the projects involved in this research proposal share this common theme. It is likely that a number of problems in cell biology which are key to normal and abnormal cell function will be answered, enhancing our understanding of renal function and disease.