Visual cue properties determine innate orientation strategy in Monarch butterflies

Animals use a wide range of environmental signals, including celestial and terrestrial cues for navigation. While celestial cues, such as the sun, play a major role in maintaining a constant heading during long-distance migration and dispersal, terrestrial cues provide an animal with a short-range navigation system, ideal to pinpoint highly specific locations. In Monarch butterflies, the simulation of a terrestrial landmark, i.e. a vertical stripe, induces an attraction behavior (all animals head toward the stimulus) while a small green light spot, simulating the sun, elicits menotactic orientation (animals adopt individual-specific headings relative to the stimulus). However, the mechanisms underlying how the animal distinguishes between a stimulus as a terrestrial landmark versus a celestial cue remains unclear. To explore this, we tested non-migratory Monarch butterflies (Danaus plexippus) in a flight simulator. The inner surface of the simulator was equipped with an array of LEDs, allowing to present different visual stimuli to the butterflies during tethered flight. By systematically manipulating the stimulus' width, height, brightness, and elevation we found that the butterflies exhibited attraction behavior to high contrast areas, like stripe edges. Menotactic behavior was not achieved by solely decreasing the stimulus to a small light spot but also required for the stimulus to be presented at higher elevation to be interpreted as a sun stimulus. These findings suggest that multiple parameters, inherently set by the butterfly's navigation system, are critical to interpret a visual stimulus as a celestial cue or terrestrial landmark, producing dynamic switches between different orientation strategies during navigation.