Regulation of Acetylcholine Receptors on Muscle

乙酰胆碱受体对肌肉的调节

• 批准号: 7925865
• 负责人: PAUL BREHM
• 金额: $ 36.07万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7925865
• 关键词: Affect; Biological Models; Cholinergic Receptors; Coupling; Data; Development; Electrical Synapse; Exhibits; Exocytosis; Failure; Feedback; Functional disorder; Funding; Genetic; Human; Ion Channel; Kinetics; Mammals; Measurement; Methodology; Molecular; Motor Neurons; Muscle; Muscle Cells; Muscle Fibers; Mutation; Nerve; Neuromuscular Diseases; Neuromuscular Junction; Nicotinic Receptors; Optical Methods; Optics; Preparation; Presynaptic Terminals; Proteins; Rana; Regulation; Reporting; Role; Shock; Signal Transduction; Skeletal Muscle; Sodium Channel; Spinal Cord; Synapses; Techniques; Testing; Toxin; Zebrafish; base; cell motility; electrical property; in vivo; mutant; patch clamp; positional cloning; postsynaptic; presynaptic; receptor; research study; response

Our studies of mutant zebrafish continue to advance understanding of synaptic transmission and inheritable forms of human neuromuscular disorders such as myasthenic syndrome, slow channel syndrome and Brody disease. The proposed studies now capitalize on mutant lines which we have shown either lack synaptic transmission altogether, or lack the ability of receptors to localize subsynaptically, the ability of muscle to release calcium, the ability of muscle to sequester calcium, or possess abnormal levels of synaptic drive suggesting miscommunication between presynaptic nerve and postsynaptic muscle. Exploiting the advantages provided by these lines, we will determine the role of synaptic activity in shaping both presynaptic and postsynaptic development. These studies will be greatly facilitated by use of our recently developed tethered toxin methodology that silences synaptic transmission in a cell autonomous fashion. Thus, we can determine the consequences of adjusting synaptic drive and/or excitability of individual cells within the motorneuron target field. Rescue of individual postsynaptic cells in mutant lines will provide a complementary approach wherein a single cell is rendered active in an otherwise silent field of target muscle. This combination of approaches will be used to determine the involvement of postsynaptic activity in the developmental loss of electrical coupling, acquisition of presynaptic transmitter release properties and establishment of postsynaptic receptor kinetics. The study is possible by virtue of our ability, for the first time in any vertebrate preparation, to simultaneously record from a motorneuron and target muscle in vivo. This is due to electrically compact muscle that allows whole cell voltage clamp and transparency, facilitating identification and patch clamp of motorneurons deep within the spinal cord. The combined advantages of new approaches and preparation now provide unique opportunities or examining long standing questions of synaptic function.

我们对突变斑马鱼的研究继续提高人们对突触传播和可继承的理解 人类神经肌肉疾病的形式，例如肌关系综合征，慢速通道综合征和Brody 疾病。拟议的研究现在利用了我们已经表明的突变管线，要么缺乏突触 完全传播，或缺乏受体次触觉定位的能力，肌肉的能力 释放钙，肌肉隔离钙的能力或具有异常水平突触驱动 暗示突触前神经和突触后肌肉之间的沟通不畅。利用 这些行提供的优点，我们将确定突触活动在塑造这两者中的作用 突触前和突触后发育。通过使用我们最近的使用，这些研究将极大地促进 开发的蛋白毒素方法论，可以以细胞自主的方式沉默突触传播。 因此，我们可以确定调节单个细胞的突触驱动和/或兴奋性的后果 在Motorneuron目标场内。在突变线中营救单个突触后细胞将提供 互补方法中，单个细胞在靶向肌肉的原本静音场中活跃。 这种方法的组合将用于确定突触后活动的参与 电耦合的发育丧失，突触前发射器释放特性的获取和 建立突触后受体动力学。这项研究可能是我们的能力，这是第一次 在任何脊椎动物制剂中，同时记录了体内肌肉和靶向肌肉。这 是由于电肌肉紧凑的肌肉，该肌肉允许全细胞电压夹和透明度，促进 脊髓深处运动神经元的识别和斑块夹。的联合优势 现在，新的方法和准备提供了独特的机会或检查长期存在的问题 突触功能。