Department of Biological Sciences, University of Delaware.
Department of Biological Sciences, University of Delaware;
J Vis Exp. 2022 Jun 7(184). doi: 10.3791/64056.
At the neuromuscular junction (NMJ), the binding of the excitatory neurotransmitter acetylcholine (ACh) to postsynaptic receptors leads to muscle contraction. As in vertebrate skeletal muscle, cholinergic signaling in the body wall muscles of the model organism Caenorhabditis elegans is required for locomotion. Exposure to levamisole, a pharmacological agonist of one class of ACh receptors on the body wall muscles, causes time-dependent paralysis of wild-type animals. Altered sensitivity to levamisole suggests defects in signaling at the NMJ or muscle function. Here, a protocol for a liquid levamisole assay performed on C. elegans grown in 24-well plates is presented. Vigorous swimming of the animals in liquid allows for the assessment and quantitation of levamisole-induced paralysis in hundreds of worms over a one-hour time period without requiring physical manipulation. This procedure can be used with both wild-type and mutants that have altered sensitivity to levamisole to demonstrate the functional consequences of altered signaling at the NMJ.
在神经肌肉接头 (NMJ) 处,兴奋性神经递质乙酰胆碱 (ACh) 与突触后受体的结合导致肌肉收缩。与脊椎动物骨骼肌一样,模型生物秀丽隐杆线虫体壁肌肉中的胆碱能信号传导对于运动是必需的。暴露于左旋咪唑(一种体壁肌肉上 ACh 受体的药理学激动剂)会导致野生型动物出现时间依赖性麻痹。对左旋咪唑的敏感性改变表明 NMJ 或肌肉功能的信号传递存在缺陷。这里介绍了一种在 24 孔板中生长的秀丽隐杆线虫上进行的液体左旋咪唑测定的方案。动物在液体中的剧烈游动允许在一个小时的时间内评估和定量数百条蠕虫中左旋咪唑诱导的麻痹,而无需进行物理操作。该程序可用于具有改变的左旋咪唑敏感性的野生型和突变体,以证明 NMJ 处改变的信号传递的功能后果。