Instituto de Investigación en Biomedicina de Buenos Aires (IBioBA-CONICET), Partner Institute of the Max Planck Society, Buenos Aires C1425FQD, Argentina.
Instituto de Investigación en Biomedicina de Buenos Aires (IBioBA-CONICET), Partner Institute of the Max Planck Society, Buenos Aires C1425FQD, Argentina
Cold Spring Harb Protoc. 2022 Aug 1;2022(8):pdb.prot107934. doi: 10.1101/pdb.prot107934.
An enormous amount of neuroscientific knowledge has been gained from studying the larval stage of From an electrophysiological point of view, the larval neuromuscular junction has played an important role in this quest for knowledge, as it presents practical advantages such as accessibility and a stereotypic pattern. The physiological properties of larval central neurons have been less explored, with information regarding mainly a few identified motoneurons available to date. This protocol describes a quick and easy dissection of the brain of wandering third-instar larvae to produce an ex vivo preparation in which central neurons can be patch-clamped. Immobilizing fresh and tiny tissues, such as larval brains, to perform successful patch-clamp recordings is a crucial step; here we explain in detail how this can be achieved using cyanoacrylate glue.
从电生理学的角度来看,幼虫的神经肌肉接头在这一知识探索中发挥了重要作用,因为它具有可及性和刻板模式等实际优势。幼虫中枢神经元的生理特性研究较少,迄今为止,有关信息主要集中在少数几个已鉴定的运动神经元上。本方案描述了一种快速简便的 wandering 三龄幼虫大脑解剖方法,可制备离体脑片,用于对中枢神经元进行膜片钳记录。固定新鲜微小的组织,如幼虫大脑,以进行成功的膜片钳记录是至关重要的一步;在这里,我们详细解释了如何使用氰基丙烯酸酯胶来实现这一点。