Abstract

Author summaryRepetitive movements such as walking, swimming, and flying are controlled by networks of neurons known as central patter generators. In many cases, the exact pattern of activity is modulated by neuropeptides, which are small signaling molecules that, unlike neurotransmitters, are broadly released within regions of the nervous system. Because of their mode of action, it can be difficult to discern the relationship between the temporal pattern of firing of peptidergic neurons and the timing of the resulting motor behavior. Here, we developed methods to analyze the patterns of activity of such weakly coupled systems as applied to ecdysis, the stereotyped sequence of behaviors used by insects to shed the remains of their old exoskeleton at the end of every molt. Key actors in this process are motoneurons (MN) and a set of neurons expressing the neuropeptide, Crustacean Cardioactive Peptide (CCAP). Combining calcium imaging, frequency analysis, computational simulations, and image processing, we determined the relationships between the activity of CCAP neurons and the resulting motor output during pupal ecdysis in the fruit fly, Drosophila melanogaster. We found that several temporal features of the activity of CCAP neurons are highly coupled to the pattern of motoneuronal activity, suggesting an active role of CCAP neurons during ecdysis. We also developed quantitative approaches that allowed us to identify a new sub-phase of ecdysis behavior.