Dissection of Drosophila Wandering Larval Brains for Patch-Clamping Neurons

An enormous amount of neuroscientific knowledge has been gained from studying the larval stage of Drosophila 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 Drosophila 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.