Increased throughput assays of locomotor dysfunction in Drosophila larvae

Larval locomotion is a sensitive readout of a range of nervous system deficits in Drosophila, and has been utilised to quantify modulation of the disease phenotype in models of human disease. Single larvae are typically analysed in series using manual quantification of parameters such as contraction rate, or grouped together and studied en-masse. Here, we describe the development of tests for the analysis of several spatially isolated third instar larvae in parallel. We rapidly quantify larval turning rate and velocity during wandering behaviour in a 4 plate assay. In a second test, larvae are recorded as they race along five parallel lanes towards a yeast stimulus. This allows increased throughput analysis of comparative genotypes simultaneously, video archiving, and detection of exacerbation or rescue of defective locomotion in a Drosophila model of tauopathy, as we demonstrate genetically and through delivery of candidate therapeutic chemicals in fly food. The tests are well-suited for rapid comparison of locomotion capability in Drosophila mutants or candidate modulation screens in Drosophila models of human disease.

► We record open-field velocity and turning rates of larvae in a 4 plate assay.
► Tau-expressing larvae show increased turning rate and lower velocity in this assay.
► Tau-expressing larvae are also slower in a novel ‘5 lane’ crawling assay.
► We detect genetic or pharmacological modulation of the tau phenotype in the tests.
► We demonstrate higher throughput chemical modulation screening with the assays.