EPITHELIAL TRANSPORT: A MORPHOLOGICAL APPROACH

上皮运输：一种形态学方法

• 批准号: 3224422
• 负责人: RICHARD M HAYS
• 金额: $ 28.99万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 1976
• 资助国家: 美国
• 起止时间: 1976-05-01 至 1994-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3224422
• 关键词: Anura; actins; angiotensins; binding proteins; bioenergetics; cell fusion; cell membrane; cryopreservation; cyclic AMP; cytoskeleton; dynein ATPase; epithelium; exocytosis; fluorescent dye /probe; hormone regulation /control mechanism; human tissue; immunocytochemistry; immunofluorescence technique; ion transport; kidney cell; laboratory rabbit; laboratory rat; membrane model; membrane permeability; membrane proteins; microtubule associated protein; protein structure function; protein transport; radioimmunoassay; radiotracer; renal tubular transport; rhodopsin; stimulant /agonist; urinary bladder; vasopressins

Two projects are described in this application. Both emphasize the relation of cell structure to epithelial transport. 1. The action of ADH: a scanning and transmission electron microscopie study. We have been able to apply current high-resolution techniques in both scanning and transmission electron microscopy (SEM and TEM) to the problem of the intracellular action of antidiuretic hormone (ADH). We will use both SEM and TEM to determine in detail the structure of the terminal web of the toad bladder epithelial cell, and the specific filamentous attachments of the cytoplasmic tubules to the luminal membrane and surrounding cytoskeleton. These tubules fuse to the membrane in response to ADH, and deliver water-conducting particles to the membrane. We have proposed a "tilting" mechanism for tubule fusion, and will seek evidence for this mechanism. We will employ tannic acid-fixed sections, resinless sections and sections coated with ultrathin metal films in our SEM and TEM sections. We will identify specific cytoskeletal components by immunocytochemistry at the light and EM level. We will isolate and characterize the water-conducting particles. We will use our techniques in a study of the mammalian kidney, emphasizing glomerular cytoskeletal structure. 2. Autoradiography of diffusible solutes. Refinements in our system of "dry" autoradiography permit the study of the transport of water-soluble solutes such as urea across epithelia, and the effects of hormones and inhibitory agents on transport. These studies will now be extended to two new problems: the paracellular transport of ions and non-electrolytes across "leaky" epithelia, and the active transport of iodide by the choroid plexus. Both projects bring new techniques to the problem of epithelial transport. There is increasing interest in fusion and exocytosis in such phenomena as neurotransmission and hormone secretion; the ADH system under investigation in this application provides one of the most clear-cut examples of the relationship of the cytoskeleton to fusion. In the human kidney, glomerular filtration appears to be greatly influenced by the contractile properties of the mesangial cell and other cellular elements surrounding the basement membrane; these may be altered in important ways in renal disease. Our autoradiographic studies show cellular transport in a new way, and add to our understanding of water and electrolyte movement.

本申请中描述了两个项目。两者都强调 细胞结构与上皮运输的关系。 1.ADH的作用：扫描和透射电子显微镜 学习。我们已经能够将当前的高分辨率技术应用于 扫描和透射电子显微镜(扫描电子显微镜和透射电子显微镜) 抗利尿激素(ADH)的细胞内作用问题。我们会 使用扫描电子显微镜和透射电子显微镜详细确定航站楼的结构 蟾蜍膀胱上皮细胞的网状结构和特定的丝状结构 细胞质小管附着在管腔膜上 围绕着细胞骨架。作为回应，这些小管融合到膜上。 到ADH，并将导水颗粒输送到膜上。我们有 提出了一种小管融合的“倾斜”机制，并将寻找证据 对于这一机制。我们将使用单宁酸固定的部分，无树脂 扫描电子显微镜和透射电子显微镜中涂覆超薄金属膜的切片和切片 横断面。我们将通过以下方式识别特定的细胞骨架组件 光镜和电子显微镜水平的免疫细胞化学。我们将隔离和 对导水颗粒进行表征。我们将使用我们的技术 哺乳动物肾脏的研究，重点是肾小球细胞骨架 结构。 2.可扩散溶质的放射自显影。对我们的系统进行改进 “干”放射自显影技术可用于研究水溶物质的运输 溶质，如尿素跨上皮细胞，以及激素和 运输中的抑制剂。这些研究现在将扩展到两个 新问题：离子和非电解质的细胞旁运输 穿过“渗漏”的上皮细胞，以及脉络膜主动转运碘 神经丛。 这两个项目都为上皮运输问题带来了新的技术。 人们对融合和胞吐现象越来越感兴趣，例如 神经传递和激素分泌；ADH系统正在研究中 在此应用程序中提供了最清晰的示例之一 细胞骨架与融合的关系。在人类肾脏中， 肾小球滤过似乎受收缩的影响很大。 肾小球系膜细胞及周围其他细胞成分的性质 基底膜；这些在肾脏中可能以重要的方式改变。 疾病。我们的放射自显影研究显示细胞在一种新的 增加了我们对水和电解质运动的理解。