Monarch butterflies (Danaus plexippus) are prominent for their annual long-distance migration from North America to their overwintering area in Central Mexico. To find their way on this long journey, they use a sun compass as their main orientation reference but will also adjust their migratory direction with respect to mountain ranges. This indicates that the migratory butterflies also attend to the panorama to guide their travels. While the compass has been studied in detail in migrating butterflies, little is known about the orientation abilities of non-migrating butterflies. Here we studied if non-migrating butterflies - that stay in a more restricted area to feed and breed - also use a similar compass system to guide their flights. Performing behavioral experiments on tethered flying butterflies in an indoor LED flight simulator, we found that the monarchs fly along straight tracks with respect to a simulated sun. When a panoramic skyline was presented as the only orientation cue, the butterflies maintained their flight direction only during short sequences suggesting that they potentially use it for flight stabilization. We further found that when we presented the two cues together, the butterflies incorporate both cues in their compass. Taken together, we here show that non-migrating monarch butterflies can combine multiple visual cues for robust orientation, an ability that may also aid them during their migration.

No abstract

Head direction can be represented in a self-centered egocentric or a viewpoint-invariant allocentric reference frame. Using the most efficient representation is especially crucial for migrating animals, like monarch butterflies (Danaus plexippus) that use the sun for orientation. With tetrode recordings from the brain of tethered flying monarch butterflies, we examined the reference frame in which insects encode heading. We show that compass neurons switch their reference frame in a state-dependent manner. In quiescence, they encode sun-bearing angles, allowing the butterfly to map the environment within an egocentric frame. However, during flight, the same neurons encode heading within an allocentric frame. This switch converts the sun from a local to a global cue, an ideal strategy for maintaining a migratory heading over large distance.

Insects are well-known for their ability to keep track of their heading direction based on a combination of skylight cues and visual landmarks. This allows them to navigate back to their nest, disperse throughout unfamiliar environments, as well as migrate over large distances between their breeding and non-breeding habitats. The monarch butterfly (Danaus plexippus) for instance is known for its annual southward migration from North America to certain trees in Central Mexico. To maintain a constant flight route, these butterflies use a time-compensated sun compass for orientation which is processed in a region in the brain, termed the central complex. However, to successfully complete their journey, the butterflies’ brain must generate a multitude of orientation strategies, allowing them to dynamically switch from sun-compass orientation to a tactic behavior toward a certain target. To study if monarch butterflies exhibit different orientation modes and if they can switch between them, we observed the orientation behavior of tethered flying butterflies in a flight simulator while presenting different visual cues to them. We found that the butterflies’ behavior depended on the presented visual stimulus. Thus, while a dark stripe was used for flight stabilization, a bright stripe was fixated by the butterflies in their frontal visual field. If we replaced a bright stripe by a simulated sun stimulus, the butterflies switched their behavior and exhibited compass orientation. Taken together, our data show that monarch butterflies rely on and switch between different orientation modes, allowing the animal to adjust orientation to its actual behavioral demands.

1Non-migrating butterflies keep directed courses when viewing a simulated sun or panoramic 2 scene. This suggest that they orient based on multiple visual cues independent of their 3 migratory context. 4 5 6 Abstract 7