SNMP1 is critical for sensitive detection of the desert locust aromatic courtship inhibition pheromone phenylacetonitrile.

BACKGROUND: Accurate detection of pheromones is crucial for chemical communication and reproduction in insects. In holometabolous flies and moths, the sensory neuron membrane protein 1 (SNMP1) is essential for detecting long-chain aliphatic pheromones by olfactory neurons. However, its function in hemimetabolous insects and its role for detecting pheromones of a different chemical nature remain elusive. Therefore, we investigated the relevance of SNMP1 for pheromone detection in a hemimetabolous insect pest of considerable economic importance, the desert locust Schistocerca gregaria, which moreover employs the aromatic pheromone phenylacetonitrile (PAN) to govern reproductive behaviors. RESULTS: Employing CRISPR/Cas-mediated gene editing, a mutant locust line lacking functional SNMP1 was established. In electroantennography experiments and single sensillum recordings, we found significantly decreased electrical responses to PAN in SNMP1-deficient (SNMP1-/-) locusts. Moreover, calcium imaging in the antennal lobe of the brain revealed a substantially reduced activation of projection neurons in SNMP1-/- individuals upon exposure to PAN, indicating that the diminished antennal responsiveness to PAN in mutants affects pheromone-evoked neuronal activity in the brain. Furthermore, in behavioral experiments, PAN-induced effects on pairing and mate choice were altered in SNMP1-/- locusts. CONCLUSIONS: Our findings emphasize the importance of SNMP1 for chemical communication in a hemimetabolous insect pest. Moreover, they show that SNMP1 plays a crucial role in pheromone detection that goes beyond long-chain aliphatic substances and includes aromatic compounds controlling reproductive behaviors.

CalciSeg: A versatile approach for unsupervised segmentation of calcium imaging data.

Recent advances in calcium imaging, including the development of fast and sensitive genetically encoded indicators, high-resolution camera chips for wide-field imaging, and resonant scanning mirrors in laser scanning microscopy, have notably improved the temporal and spatial resolution of functional imaging analysis. Nonetheless, the variability of imaging approaches and brain structures challenges the development of versatile and reliable segmentation methods. Standard techniques, such as manual selection of regions of interest or machine learning solutions, often fall short due to either user bias, non-transferability among systems, or computational demand. To overcome these issues, we developed CalciSeg, a data-driven and reproducible approach for unsupervised functional calcium imaging data segmentation. CalciSeg addresses the challenges associated with brain structure variability and user bias by offering a computationally efficient solution for automatic image segmentation based on two parameters: regions' size limits and number of refinement iterations. We evaluated CalciSeg efficacy on datasets of varied complexity, different insect species (locusts, bees, and cockroaches), and imaging systems (wide-field, confocal, and multiphoton), showing the robustness and generality of our approach. Finally, the user-friendly nature and open-source availability of CalciSeg facilitate the integration of this algorithm into existing analysis pipelines.

An effective hydrodynamic description of marching locusts.

A fundamental question in complex systems is how to relate interactions between individual components ('microscopic description') to the global properties of the system ('macroscopic description'). Furthermore, it is unclear whether such a macroscopic description exists and if such a description can capture large-scale properties. Here, we address the validity of a macroscopic description of a complex biological system using the collective motion of desert locusts as a canonical example. One of the world's most devastating insect plagues begins when flightless juvenile locusts form 'marching bands'. These bands display remarkable coordinated motion, moving through semiarid habitats in search of food. We investigated how well macroscopic physical models can describe the flow of locusts within a band. For this, we filmed locusts within marching bands during an outbreak in Kenya and automatically tracked all individuals passing through the camera frame. We first analyzed the spatial topology of nearest neighbors and found individuals to be isotropically distributed. Despite this apparent randomness, a local order was observed in regions of high density in the radial distribution function, akin to an ordered fluid. Furthermore, reconstructing individual locust trajectories revealed a highly aligned movement, consistent with the one-dimensional version of the Toner-Tu equations, a generalization of the Navier-Stokes equations for fluids, used to describe the equivalent macroscopic fluid properties of active particles. Using this effective Toner-Tu equation, which relates the gradient of the pressure to the acceleration, we show that the effective 'pressure' of locusts increases as a linear function of density in segments with the highest polarization (for which the one-dimensional approximation is most appropriate). Our study thus demonstrates an effective hydrodynamic description of flow dynamics in plague locust swarms.

Active smelling in the American cockroach.

Motion plays an essential role in sensory acquisition. From changing the position in which information can be acquired to fine-scale probing and active sensing, animals actively control the way they interact with the environment. In olfaction, movement impacts the time and location of odour sampling as well as the flow of odour molecules around the olfactory organs. Employing a detailed spatiotemporal analysis, we investigated how insect antennae interact with the olfactory environment in a species with a well-studied olfactory system - the American cockroach. Cockroaches were tested in a wind-tunnel setup during the presentation of odours with different attractivity levels: colony extract, butanol and linalool. Our analysis revealed significant changes in antennal kinematics when odours were presented, including a shift towards the stream position, an increase in vertical movement and high-frequency local oscillations. Nevertheless, the antennal shifting occurred predominantly in a single antenna while the overall range covered by both antennae was maintained throughout. These findings hold true for both static and moving stimuli and were more pronounced for attractive odours. Furthermore, we found that upon odour encounter, there was an increase in the occurrence of high-frequency antennal sweeps and vertical strokes, which were shown to impact the olfactory environment's statistics directly. Our study lays out a tractable system for exploring the tight coupling between sensing and movement, in which antennal sweeps, in parallel to mammalian sniffing, are actively involved in facilitating odour capture and transport, generating odour intermittency in environments with low air movement where cockroaches dwell.

Coding of odour and space in the hemimetabolous insect Periplaneta americana.

The general architecture of the olfactory system is highly conserved from insects to humans, but neuroanatomical and physiological differences can be observed across species. The American cockroach, inhabiting dark shelters with a rather stable olfactory landscape, is equipped with long antennae used for sampling the surrounding air-space for orientation and navigation. The antennae's exceptional length provides a wide spatial working range for odour detection; however, it is still largely unknown whether and how this is also used for mapping the structure of the olfactory environment. By selectively labelling antennal lobe projection neurons with a calcium-sensitive dye, we investigated the logic of olfactory coding in this hemimetabolous insect. We show that odour responses are stimulus specific and concentration dependent, and that structurally related odorants evoke physiologically similar responses. By using spatially confined stimuli, we show that proximal stimulations induce stronger and faster responses than distal ones. Spatially confined stimuli of the female pheromone periplanone B activate a subregion of the male macroglomerulus. Thus, we report that the combinatorial logic of odour coding deduced from holometabolous insects applies also to this hemimetabolous species. Furthermore, a fast decrease in sensitivity along the antenna, not supported by a proportionate decrease in sensillar density, suggests a neural architecture that strongly emphasizes neuronal inputs from the proximal portion of the antenna.

Intersegmental coupling and recovery from perturbations in freely running cockroaches.

Cockroaches are remarkably stable runners, exhibiting rapid recovery from external perturbations. To uncover the mechanisms behind this important behavioral trait, we recorded leg kinematics of freely running animals in both undisturbed and perturbed trials. Functional coupling underlying inter-leg coordination was monitored before and during localized perturbations, which were applied to single legs via magnetic impulses. The resulting transient effects on all legs and the recovery times to normal pre-perturbation kinematics were studied. We estimated coupling architecture and strength by fitting experimental data to a six-leg-unit phase oscillator model. Using maximum-likelihood techniques, we found that a network with nearest-neighbor inter-leg coupling best fitted the data and that, although coupling strengths vary among preparations, the overall inputs entering each leg are approximately balanced and consistent. Simulations of models with different coupling strengths encountering perturbations suggest that the coupling schemes estimated from our experiments allow animals relatively fast and uniform recoveries from perturbations.

Synergistic olfactory processing for social plasticity in desert locusts.

Desert locust plagues threaten the food security of millions. Central to their formation is crowding-induced plasticity, with social phenotypes changing from cryptic (solitarious) to swarming (gregarious). Here, we elucidate the implications of this transition on foraging decisions and corresponding neural circuits. We use behavioral experiments and Bayesian modeling to decompose the multi-modal facets of foraging, revealing olfactory social cues as critical. To this end, we investigate how corresponding odors are encoded in the locust olfactory system using in-vivo calcium imaging. We discover crowding-dependent synergistic interactions between food-related and social odors distributed across stable combinatorial response maps. The observed synergy was specific to the gregarious phase and manifested in distinct odor response motifs. Our results suggest a crowding-induced modulation of the locust olfactory system that enhances food detection in swarms. Overall, we demonstrate how linking sensory adaptations to behaviorally relevant tasks can improve our understanding of social modulation in non-model organisms.

The chemical ecology of locust cannibalism.

An anticannibalistic signaling pathway offers a new understanding of locust swarm formation.

Information integration for decision-making in desert locusts.

Locust swarms can extend over hundred kilometers, and starvation compels this ancient pest to devour everything in its path. Theory suggests that gregarious behavior benefits foraging efficiency, yet the role of social cohesion remains elusive. To this end, we collected high-resolution trajectories of individual and grouped gregarious desert locusts in a 2-choice assay with patches of either similar or different quality. Carefully maintaining animals' identities allowed us to monitor each individual's experience and estimate the leaky accumulation process of personally acquired and socially derived evidence. We fitted data to a Bayesian model to gain insight into the decision-making system for patch selection. By disentangling the relative contribution of each information class, our study suggests that locusts balance incongruent evidence but reinforce congruent ones. We provide insight into the collective foraging decisions of social (but non-eusocial) insects and present locusts as a powerful empirical system to study individual choices and consequent collective dynamics.

The social brain of 'non-eusocial' insects.

Decisions are seldom entirely devoid of social influence. Even in organisms that have traditionally been considered non-social, the social environment plays an important role in mediating behavior. Here we review the current knowledge regarding the neural basis of social behaviors in non-eusocial insects, with a particular focus on fruit flies, cockroaches and locusts. Each are shown to offer valuable, and complementary, insights into how social behavior is mediated at the neural level. The presented studies demonstrate that social cues, which are integrated in primary sensory areas, exert a considerable influence on behavior. Further studies with these models, and others, will provide important insights into the diversity of social behaviors, and into the way that these are encoded in dedicated brain and neuronal structures.

Social modulation of individual preferences in cockroaches.

In social species, decision-making is both influenced by, and in turn influences, the social context. This reciprocal feedback introduces coupling across scales, from the neural basis of sensing, to individual and collective decision-making. Here, we adopt an integrative approach investigating decision-making in dynamical social contexts. When choosing shelters, isolated cockroaches prefer vanillin-scented (food-associated) shelters over unscented ones, yet in groups, this preference is inverted. We demonstrate that this inversion can be replicated by replacing the full social context with social odors: presented alone food and social odors are attractive, yet when presented as a mixture they are avoided. Via antennal lobe calcium imaging, we show that neural activity in vanillin-responsive regions reduces as social odor concentration increases. Thus, we suggest that the mixture is evaluated as a distinct olfactory object with opposite valence, providing a mechanism that would naturally result in individuals avoiding what they perceive as recently exploited resources.

The subesophageal ganglion modulates locust inter-leg sensory-motor interactions via contralateral pathways.

The neural control of insect locomotion is distributed among various body segments. Local pattern-generating circuits at the thoracic ganglia interact with incoming sensory signals and central descending commands from the head ganglia. The evidence from different insect preparations suggests that the subesophageal ganglion (SEG) may play an important role in locomotion-related tasks. In a previous study, we demonstrated that the locust SEG modulates the coupling pattern between segmental leg CPGs in the absence of sensory feedback. Here, we investigated its role in processing and transmitting sensory information to the leg motor centers and mapped the major related neural pathways. Specifically, the intra- and inter-segmental transfer of leg-feedback were studied by simultaneously monitoring motor responses and descending signals from the SEG. Our findings reveal a crucial role of the SEG in the transfer of intersegmental, but not intrasegmental, signals. Additional lesion experiments, in which the intersegmental connectives were cut at different locations, together with double nerve staining, indicated that sensory signals are mainly transferred to the SEG via the connective contralateral to the stimulated leg. We therefore suggest that, similar to data reported for vertebrates, insect leg sensory-motor loops comprise contralateral ascending pathways to the head and ipsilateral descending ones.