ULTRASTRUCTURE AT ACH RECEPTOR CLUSTERS IN MUSCLE

肌肉中每个受体簇的超微结构

• 批准号: 3396087
• 负责人: ROBERT W SEALOCK
• 金额: $ 10.16万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 1979
• 资助国家: 美国
• 起止时间: 1979-04-01 至 1990-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3396087
• 关键词: acetylcholine; acetylcholinesterase; basolateral membrane; cell cell interaction; cell membrane; cytoplasm; cytoskeleton; electron microscopy; freeze etching; histochemistry /cytochemistry; immunochemistry; immunofluorescence technique; neuromuscular junction; saltwater environment; sarcolemma; striated muscles; synapses

This project is an outgrowth of work in this laboratory using ultrastructural methods and immunocytochemistry to study cytoplasmic proteins associated with the postsynaptic membrane of Torpedo electric tissue. The main long-term goal has been, and remains, to identify the mechanisms by which accumulations ("clusters") of acetylcholine receptor molecules are stabilized ("anchored") against lateral diffusion in the plasma membrane. In this proposal, the underlying idea--that anchoring is mediated by cytoskeletal and other cytoplasmic structures--is expanded to include a role for cluster-specific basal lamina, and the work is shifted to skeletal muscle from rat and Xenopus laevis. The previous methods plus freeze-fracture/deep-etch electron microscopy will be applied to clusters on muscle cells in culture, developing neuromuscular junctions, and the mature junction. Cytoskeletal, other cytoplasmic, and extracellular proteins at clusters will be identified immunocytochemically and localized relative to the receptor, the structures in which these proteins occur will be visualized in 3-dimensions, and how identified proteins fit into these structures will be determined. It is hoped that by judicious choices of sample and with the aid of cell biological manipulation of clusters, the various roles of these structures in receptor anchoring, in maintenance of the cell-cell contact nature of the junction, and in other possible synaptic activities can be delineated.

该项目是该实验室的工作的产物 超微结构方法和免疫细胞化学研究细胞质 与鱼雷电气的突触后膜相关的蛋白质 组织。 长期的主要目标是并且仍然是确定 乙酰胆碱受体积累（“簇”）的机制 分子稳定（“锚定”）针对横向扩散 质膜。 在这个建议中，基本的想法 - 锚定是 由细胞骨架和其他细胞质结构介导的 包括特定于集群的基底层薄片的作用，工作被转移 从大鼠和爪蟾骨肌肉到骨骼肌。 以前的方法加上 冻结裂缝/深度蚀刻电子显微镜将应用于簇 在培养中的肌肉细胞上，发展神经肌肉连接和 成熟的交界处。 细胞骨骼，其他细胞质和细胞外 簇的蛋白质将被鉴定出免疫细胞化学和局部 相对于受体，发生这些蛋白质的结构将 在三维中可视化，以及如何鉴定的蛋白质适合这些 结构将被确定。 希望通过明智的选择 样品并借助细胞生物学操纵簇， 这些结构在受体锚定中的各种作用，维持 连接点的细胞电池接触性质，其他可能 可以划定突触活动。