Dissection of First- and Second-Instar Drosophila Larvae for Electrophysiological Recording from Neurons: The Flat (or Fillet) Preparation

INTRODUCTION

The fruit fly Drosophila melanogaster has been instrumental in expanding our understanding of early aspects of neural development. The use of this model system has greatly added to our knowledge of neural cell-fate determination, axon guidance, and synapse formation. It has also become possible to access and make electrophysiological recordings directly from neurons in situ in an intact central nervous system (CNS), which has facilitated studies of the development and regulation of neuronal signaling. It is possible to obtain electrophysiological recordings from all stages of Drosophila. Exposure of the intact Drosophila CNS is a prerequisite for such electrophysiological recordings. The dissection procedure described here can be applied to both late-stage embryos (stage 16 onward) and larvae. Because of their size, third-instar larvae are more difficult to flatten using this method and, if recording from this stage, the reader might consider using insect pins for the dissection or isolating the CNS using an alternative method. The dissection should take <10 min if all preparation work has been completed in advance. Owing to the short life span of the dissected larva, it is not recommended that the procedure be stopped or the preparation stored for later use.