Transcuticular optical imaging of stimulus-evoked neural activities in the Drosophila peripheral nervous system

The nervous system of Drosophila is widely used to study neuronal signal processing because the activities of neurons can be controlled and monitored by cell type-specific expression of genetically encoded actuator and sensor proteins. Measuring neural activities in adult flies, however, usually requires surgical approaches to penetrate the firm and pigmented cuticular exoskeleton. Interfering with this exoskeleton is critical in the case of the peripheral nervous system (PNS), as sensory neurons are often located directly beneath the cuticle and are associated with specialized stimulus-receiving and -conducting cuticular structures. In this article, we describe how the activities of these neurons can be probed nondestructively through the cuticle if a genetically encoded fluorescent protein sensor with strong baseline fluorescence is used. The method is exemplified for mechanosensory neurons in the adult antenna but can also be applied to many other PNS neurons, as is shown for the femoral chordotonal organ located in the fly's leg.