Cold stress results in sustained locomotor and behavioral deficits in Drosophila melanogaster

Tolerance of climatic stressors is an important predictor of the current distribution of insect species, their potential to invade new environments, and their responses to rapid climate change. Cold stress causes acute injury to nerves and muscles, and here we tested the hypothesis that low temperature causes sublethal deficits in locomotor behaviors that are dependent on neuromuscular function. To do so, we applied a previously developed assay, the rapid iterative negative geotaxis (RING) assay, to investigate behavioral consequences of cold stress in Drosophila melanogaster. The RING assay allows for rapid assessment of negative geotaxis behavior by quantifying climbing height and willingness to climb after cold stress. We exposed flies to cold stress at 0°C and assessed the extent to which duration of cold stress, recovery time, and cold acclimation influenced climbing performance. There was a clear dose-response relationship between cold exposure and performance deficits, with climbing height and willingness decreasing as cold exposure increased from 2 to 24 hr. Following cold exposure of an intermediate duration (12 hr), climbing height and willingness gradually improved as recovery time increased from 4 to 72 hr but flies never fully recovered. Finally, cold acclimation improved overall climbing height and willingness in both untreated and cold-stressed flies but did not prevent a reduction in climbing performance. Thus, cold stress causes deficits in locomotor and behavior that are dependent on the dose of cold exposure and persist long after the stress subsides. These results likely have implications for the ecological and evolutionary responses of insect populations to thermally variable environments.