Drosophila require both green and UV wavelengths for sun orientation but lack a time-compensated sun compass

Celestial orientation and navigation are performed by many organisms in contexts as diverse as migration, nest finding and straight-line orientation. The vinegar fly, Drosophila melanogaster, performs menotaxis in response to celestial cues during tethered flight and can disperse more than 10 km under field conditions. However, we still do not understand how spectral components of celestial cues and pauses in flight impact heading direction in flies. To assess individual heading, we began by testing flies in a rotating tether arena using a single green LED as a stimulus. We found that flies robustly perform menotaxis and fly straight for at least 20 min. Flies maintain their preferred heading directions after experiencing a period of darkness or stopping flight, even up to 2 h, but reset their heading when the LED changes position, suggesting that flies do not treat this stimulus as the sun. Next, we assessed the flies' responses to a UV spot alone or a paired UV-green stimulus - two dots situated 180 deg apart to simulate the solar and antisolar hemispheres. We found that flies respond to UV much as they do to green light; however, when the stimuli are paired, flies adjust for sudden 90 deg movements, performing sun orientation. Lastly, we found no evidence of a time-compensated sun compass when we moved the paired stimuli at 15 deg h-1 for 6 h. This study demonstrates that wavelength influences how flies respond to visual cues during flight, shaping the interpretation of visual information to execute an appropriate behavioral response.