DOI: 10.5061/dryad.pzgmsbctg
title: Failure to mate enhances investment in behaviors that may promote mating reward and impairs the ability to cope with stressors via a subpopulation of Neuropeptide F receptor neurons

description: Living in dynamic environments such as the social domain, where
 interaction with others determines the reproductive success of
 individuals, requires the ability to recognize opportunities to obtain
 natural rewards and cope with challenges that are associated with
 achieving them. As such, actions that promote survival and reproduction
 are reinforced by the brain reward system, whereas coping with the
 challenges associated with obtaining these rewards is mediated by
 stress-response pathways, the activation of which can impair health and
 shorten lifespan. While much research has been devoted to understanding
 mechanisms underlying the way by which natural rewards are processed by
 the reward system, less attention has been given to the consequences of
 failure to obtain a desirable reward. As a model system to study the
 impact of failure to obtain a natural reward, we used the well-established
 courtship suppression paradigm in Drosophila melanogaster as means to
 induce repeated failures to obtain sexual reward in male flies. We
 discovered that beyond the known reduction in courtship actions caused by
 interaction with non-receptive females, repeated failures to mate induce a
 stress response characterized by persistent motivation to obtain the
 sexual reward, reduced male-male social interaction, and enhanced
 aggression. This frustrative-like state caused by the conflict between
 high motivation to obtain sexual reward and the inability to fulfill their
 mating drive impairs the capacity of rejected males to tolerate stressors
 such as starvation and oxidative stress. We further show that sensitivity
 to starvation and enhanced social arousal is mediated by the disinhibition
 of a small population of neurons that express receptors for the fly
 homologue of neuropeptide Y. Our findings demonstrate for the first time
 the existence of social stress in flies and offers a framework to study
 mechanisms underlying the crosstalk between reward, stress, and
 reproduction in a simple nervous system that is highly amenable to genetic
 manipulation.
description: This dataset includes all the experiments done in this particular
 manuscript and include: FlyBowl raw data, QPCR, aggression, Mating
 duration, sensitivity to stress curves, activity monitor and
 metabolomics
description: # Failure to mate enhances investment in behaviors that may promote mating
 reward and impairs the ability to cope with stressors via a subpopulation
 of Neuropeptide F receptor neurons.
 Give a brief summary of
 dataset contents, contextualized in experimental procedures and results.
 ## Description of the data and file structure This dataset contains all
 the raw data for this particular manuscript. It includes per-fly and
 per-frame data for the experiments done using the Flybowl system, QPCR
 data, metabolomic data, aggression, sleep, mating and courtship
 suppression data. The file name describes the relevant figure and
 genotype. Empty cells contain n/a. **Below is the list of the files:** 1.
 Courtship suppression data-set relevant for Figure S3. The table contains
 manual scoring of various courtship related actions (number of events and
 duration). This includes latency to first courtship action (sec), overall
 time spend courting (sec), latency to first attempt of copulation (sec),
 number of coupulation attempts, total length of wing vibration actions
 (sec), grooming events, nubmer of licking events. Each action is depicted
 in a separate sheet. 2. Metabolomic raw data relevant for Figure S5. The
 table contains raw readings of LC-MS. 3. Starvation resistance raw data
 relevant for figures 4, 5 and S6-8. This table contains the counts of live
 flies along starvation resistance assays. 4. Copulation duration raw data
 relevant for Figure 3D. The table depicts the overall length of copulation
 events in seconds. 5. Aggression raw data relevant for Figure 3A. depics
 counts of lunges actions. 6. Aggression (mixed pairs) raw data relevant
 for Figure 3B-C. depicts counts of lunges events. 7. Raw data for glucose,
 weight, and TAG relevant for Figure S5A-D. contains fluoremetric measures
 of glucose and TAG as well as total weight of the flies. 8. Sleep raw data
 relevant for Figure S4 B-D. This table contains raw data for movement
 activity of single flies, and calculations of events in which they
 displayed sleep. 9. QPCR raw data for four repeats: Figure 3E. This figure
 is based on 4 biological repeats of qPCR analysis (Biorad). Each table
 contains the raw CT values of the measured transcripts. 10. Fly Bowl
 experiments raw data contain per fly data of the following features: |
 **Definition** | **Description** | |
 :--------------------------------------------------------- |
 :------------------------------------------------------------------------------------------------------------------ | | Walk | Fly moves. | | Stop | Fly is still. | | Turn | Changes in fly’s direction. | | Touch | Fly actively touches another fly. | | Social cluster | Fly sits in a group of 3 or more flies. | | Long distance Approach | Fly approaches from a distance another fly and perform interaction. | | Short distance approach | Fly performs an interaction during when in social cluster. | | Grooming | Fly grooms. | | Chase | Fly chases another fly. | | Chain | Chase with 3 or more flies. | | Song | Fly moves one wing next to another fly. | | Behavior bout length | Length of the longest sequence of frames in which the behavior occurred per fly. | | Behavior inter bout length | Duration between bouts of same behavior. | | dnose2ell | Minimum distance from any point of this animal nose to the ellipse of other flies. | | absanglefrom1to2nose2ell | Absolute difference between direction to closest animal and current animal’s orientation (rad). | | absdtheta | Angular speed (rad/s). | | absphidiff anglesub | Absolute difference in velocity direction between current animal and closest animal(rad). | | absphidiff nose2ell | Absolute difference in velocity direction between current animal and closest animal (rad). | | absthetadiff anglesub | Absolute difference in orientation between current animal and closest animal (rad). | | absthetadiff nose2ell | Absolute difference in orientation between this animal and closest animal (rad). | | anglefrom1to2 anglesub | Angle to closest animal’s centroid in current animal’s coordinate system (rad). | | anglefrom1to2 nose2ell | Angle to closest animal’s centroid in current animal’s coordinate system (rad). | | angleonclosestfly | Angle of the current animal’s centroid in the closest animal’s coordinate system (rad). | | anglesub | Maximum total angle of animal’s field of view (fov) occluded by another animal (rad). | | danglesub | Change in maximum total angle of animal’s view occluded by another animal (rad/s). | | dcenter | Minimum distance from this animal’s center to other animal’s center (mm). | | ddcenter | Change in minimum distance between this animal’s center and other flies’ centers (mm/s). | | dist2wall | Distance to the arena wall from the animal’s center (mm). | | dphi | Change in the velocity direction (rad/s). | | dtheta | Angular velocity (rad/s). | | nflies\_close | Number of flies within 2 body lengths (4a). | | velmag | Speed of the center of rotation (mm/s). | | Density SD by length of interactions (LOI) | Accumulated interactions’ length relative to the maximum interactions’ length possible. | | Modularity by length of interactions (LOI) | Representation of how much the network is divided into modules according to interactions’ length. | | Strength by length of interactions (LOI) | Length of interactions of a certain fly. | | SD Strength according to length of interactions (LOI) | Standard divination of the strengths according to interactions’ length of flies from the same movie. | | Betweenness Centrality by length of interactions (LOI) | A measure of centrality of a certain fly based on shortest paths according to interactions’ length. | | SD Betweenness Centrality by length of interactions (LOI) | Standard divination of the betweenness centralities according to interactions’ length of flies from the same movie. | | Density by number of interactions (NOI) | Interactions’ number relative to the maximum interactions’ number possible. | | Modularity Strength by number of interactions (NOI) | Representation of how much the network is divided into modules according to interactions’ number. | | Strength by number of interactions (NOI) | Number of interactions of a certain fly. | | SD Strength by number of interactions (NOI) | Standard divination of the strengths according to interactions’ number of flies from the same movie. | | Betweenness Centrality by number of interactions (NOI) | A measure of centrality of a certain fly based on shortest paths according to interactions’ number. | | SD Betweenness centrality (by number of interactions (NOI) | Variance of the betweenness centralities according to interactions’ number of flies from the same movie. | The same feature are depicted in the following panels:  Rejected, Naïve, Mated Figures 1, 2 and S2  Activation of all NPFR neurons Figure S6B  Activation of Dh44 neurons Figure S7A  Activation of NPFR-NPF neurons Figure S6C Activation of NPFR-TK neurons Figure S8C Per fly, Per frame data: Rejected, naïve and mated relevant for Figure 2F. activation of NPFR_TK neurons relevant for Figure 5E-I