Title: Serotonergic Modulation of Walking Behavior in Drosophila melanogaster
Description: Walking is an essential behavior across the animal kingdom. To navigate complex environments, animals must have highly robust, yet flexible locomotor behaviors. One crucial aspect of this process is the selection of an appropriate walking speed. Speed shifts entail not only the scaling of behavioral parameters (such as faster steps) but also changes in coordination to produce different gaits, and the details of how this switch occurs are currently unknown. Modulatory substances, particularly small biogenic amine neurotransmitters, can alter the output and even the connectivity of motor circuits. This work addresses the hypothesis that one such neuromodulator – serotonin (5HT) – is a key regulator of walking speed at the level of motor circuitry. To explore this question, I use the model organism Drosophila melanogaster which, like vertebrates, displays complex coordinated locomotion at a wide range of speeds. In Chapter 2, I will describe our efforts to characterize the anatomy of the serotonergic cell populations that provide direct input to motor circuitry.

I find that innervation of the neuropil of the ventral nerve cord - a structure roughly analagous to the mammalian spinal cord - is provided primarily by local modulatory interneurons. Using stochastic single cell labeling techniques, I will detail the specific anatomy of individual neuromodulatory cells, and also the distribution of synapses across their processes. In Chapter 3, I will show that optogenetic activation or tonic inhibition of VNC serotonergic neurons produces opposing shifts in walking speed. To analyze behavior, I will use two complementary approaches. On the one hand, I will use an arena assay to holistically assess walking velocity and frequency. On the other, I will use a behavioral assay developed in the lab - the Flywalker - to assess walking kinematics at high resolution. The combination of these technique will give us a broad and specific picture of how the VNC serotonergic system modulates walking. In Chapter 4, I will identify natural behavioral contexts under which serotonin is used to shift walking behavior.

I will use a variety of paradigms that induce animals to shift their speed, from changes in orientation and nutrition state, to pulses of light, odor, and a vibration. I will assess the requirement for the VNC serotonergic system under all of these conditions, to build a clearer picture of its role in modulating behavioral adaptation. In Chapter 5, I will describe our efforts, in collaboration with Pavan Ramdya's lab at EPFL, to functionally image VNC serotonergic cells while the animal is walking, to understand how activity is endogenously regulated in this population. Finally, in Chapter 6 I will characterize the circuit elements which might be responsible for serotonin's effect on walking. I will use recently developed mutant lines to identify the particular serotonergic receptors responsible for enacting shifts in walking behavior. Using genetic labeling tools, I will identify potential targets of serotonergic signaling in the VNC, and formulate a model by which action on these targets could adjust locomotor output.

Altogether, this work seeks to characterize the anatomy and behavioral role of the VNC serotonergic system in Drosophila. I hope that through this work, I will gain a deeper understanding of not only this particular modulatory system in this particular behavioral context, but also of how static circuits are conferred with essential flexibility in behaving animals.