Extracting temporal relationships between weakly coupled peptidergic and motoneuronal signaling: application toDrosophilaecdysis behavior
Authors: Piñeiro Miguel , Mena Wilson , Ewer John , Orio Patricio 
Published: April 2021

AbstractNeuromodulators, such as neuropeptides, can regulate and reconfigure neural circuits to alter their output, affecting in this way animal physiology and behavior. The interplay between the activity of neuronal circuits, their modulation by neuropeptides, and the resulting behavior, is still poorly understood. Here, we present a quantitative framework to study the relationships between the temporal pattern of activity of peptidergic neurons and of motoneurons duringDrosophilaecdysis behavior, a highly stereotyped motor sequence that is critical for insect growth. We analyzed, in the time and frequency domains, simultaneous intracellular calcium recordings of peptidergic CCAP (crustacean cardioactive peptide) neurons and motoneurons obtained from isolated central nervous systems throughout fictive ecdysis behavior inducedex vivoby Ecdysis triggering hormone. We found that the activity of both neuronal populations is tightly coupled in a cross-frequency manner, suggesting that CCAP neurons modulate the faster oscillation of motoneurons. To explore this idea further, we used a probabilistic logistic model to show that calcium dynamics in CCAP neurons can predict the oscillation of motoneurons, both in a simple model and in a conductance-base model capable of simulating many of the observed neural dynamics features. Finally, we developed an algorithm to quantify the motor behavior observed in videos of pupal ecdysis, and compared their features to the patterns of neuronal calcium activity recordedex vivo. We found that the motor activity of the intact animal is more regular than the motoneuronal activity recorded from theex vivopreparations during fictive ecdysis behavior; the analysis of movement patterns also allowed us to identify a new post-ecdysis phase.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 this 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 real-time 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 patter of activity motoneuronal pattern, suggesting an active role of CCAP neurons during ecdysis. We also developed quantitative approaches that allowed us to identify a new ecdysis sub-phase.