FORMATION AND ELIMINATION OF SYNAPSES

突触的形成和消除

• 批准号: 2263316
• 负责人: CHIEN-PING KO
• 金额: $ 14.22万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 1981
• 资助国家: 美国
• 起止时间: 1981-12-01 至 1996-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2263316
• 关键词: Rana; Schwann cells; Torpedo; Xenopus; affinity chromatography; alternatives to animals in research; antibody; binding proteins; cholinergic receptors; embryo /fetus tissue /cell culture; extracellular matrix; fish electric organ; fluorescent dye /probe; gel electrophoresis; immunocytochemistry; innervation; laboratory rabbit; lectin; molecular weight; nerve endings; neural degeneration; neural plasticity; neurogenesis; neuromuscular junction; stainings; synaptogenesis; western blottings

The long-term objectives of this research are to elucidate the cellular and molecular mechanisms of development, maintenance and plasticity of the synapse. The present proposal will focus on the involvement of extracellular matrix (ECM) glycoconjugates in these processes at the neuromuscular junction (NMJ) using peanut agglutinin (PNA) as a molecular probe. The specific aims are: (I). To test the hypothesis that expression of synaptic ECM precedes the nerve terminal outgrowth during synaptic remodeling. (A) Identified NMJs will be observed repeatedly in vivo with video-enhanced microscopy followed by electron microscopy. In addition, whether Schwann cell processes lead the nerve terminal will be studied. (B) The dynamic relationship among nerve terminals, synaptic ECM and acetylcholine receptors during the active growth of sprouting will be examined in vivo. (II). To characterize PNA-binding molecules (PNA-BMs) and test their functions. (A) Affinity chromatography, gel electrophoresis and lectin blotting will be used to isolate and identify the PNA-BMs. (B) Antibodies against PNA-BMs will be produced and immunocytochemistry and immunoblotting performed. (C) The effects of affinity-purified PNA-BMs and functional perturbation with antibodies on neurite outgrowth in vitro will be examined. (D) Antibodies will be applied to perturb the function of PNA-BMs in synapse formation and maintenance in vitro and in vivo. The proposed research would provide novel concepts on the role of the ECM and Schwann cells during synaptic remodeling in living adult animals. The proposal would also characterize new synapse-specific ECM molecules and test the function in formation, plasticity and maintenance of the NMJ. The result may lead to a better understanding of neuromuscular diseases and restoration after trauma. The study may also provide new thinking on learning and memory, which may involve remodeling of synaptic connections in the brain.

这项研究的长期目标是阐明细胞 以及发育、维持和可塑性的分子机制 突触。 本提案将重点关注 细胞外基质（ECM）糖复合物在这些过程中 使用花生凝集素 (PNA) 作为分子的神经肌肉接头 (NMJ) 探测。 具体目标是： （我）。检验突触 ECM 表达先于突触 ECM 表达的假设 突触重塑过程中神经末梢的生长。 (A) 已确定的 NMJ 将通过视频增强显微镜在体内反复观察 其次是电子显微镜。 另外，雪旺细胞是否 将研究导致神经末梢的过程。 （二）动态 神经末梢、突触ECM和乙酰胆碱之间的关系 将在体内检查发芽活跃生长期间的受体。 （二）。表征 PNA 结合分子 (PNA-BM) 并测试其 功能。 (A) 亲和层析、凝胶电泳和凝集素 印迹法将用于分离和鉴定 PNA-BM。 (二) 将产生针对 PNA-BM 的抗体，并进行免疫细胞化学和 进行免疫印迹。 (C) 亲和纯化的 PNA-BM 的作用 以及抗体对体外神经突生长的功能干扰 将接受检查。 (D) 将应用抗体来扰乱功能 PNA-BM 在体外和体内突触形成和维持中的作用。 拟议的研究将为 ECM 的作用提供新概念 和雪旺细胞在活体成年动物突触重塑过程中的作用。 该提案还将描述新的突触特异性 ECM 分子的特征 并测试其形成、可塑性和维持功能 NMJ。 该结果可能有助于更好地理解神经肌肉 疾病和创伤后的恢复。 该研究还可能提供新的 对学习和记忆的思考，这可能涉及突触的重塑 大脑中的连接。