Intergenerational genotypic interactions drive collective behavioural cycles in a social insect.

Many social animals display collective activity cycles based on synchronous behavioural oscillations across group members. A classic example is the colony cycle of army ants, where thousands of individuals undergo stereotypical biphasic behavioural cycles of about one month. Cycle phases coincide with brood developmental stages, but the regulation of this cycle is otherwise poorly understood. Here, we probe the regulation of cycle duration through interactions between brood and workers in an experimentally amenable army ant relative, the clonal raider ant. We first establish that cycle length varies across clonal lineages using long-term monitoring data. We then investigate the putative sources and impacts of this variation in a cross-fostering experiment with four lineages combining developmental, morphological and automated behavioural tracking analyses. We show that cycle length variation stems from variation in the duration of the larval developmental stage, and that this stage can be prolonged not only by the clonal lineage of brood (direct genetic effects), but also of the workers (indirect genetic effects). We find similar indirect effects of worker line on brood adult size and, conversely (but more surprisingly), indirect genetic effects of the brood on worker behaviour (walking speed and time spent in the nest).

Temporal and structural neural asymmetries in insects.

Neural asymmetries of the bilateral parts of the nervous system are found throughout the animal kingdom. The relative low complexity and experimental accessibility of the insect nervous system makes it well suited for studying the functions of neural asymmetries and their underlying mechanisms. Recent findings in insects reveal hardwired asymmetries in their peripheral and central nervous systems, which affect sensory perception, motor behaviours and cognitive-related tasks. Together, these findings underscore the tendency of the nervous system to segregate between the activities of its right and left sides either transiently or as permanent lateralized specializations.

Collective motion as a distinct behavioral state of the individual.

The collective motion of swarms depends on adaptations at the individual level. We explored these and their effects on swarm formation and maintenance in locusts. The walking kinematics of individual insects were monitored under laboratory settings, before, as well as during collective motion in a group, and again after separation from the group. It was found that taking part in collective motion induced in the individual unique behavioral kinematics, suggesting the existence of a distinct behavioral mode that we term a "collective-motion-state." This state, characterized by behavioral adaptation to the social context, is long lasting, not induced by crowding per se, but only by experiencing collective motion. Utilizing computational models, we show that this adaptability increases the robustness of the swarm. Overall, our findings suggest that collective motion is not only an emergent property of the group but also depends on a behavioral mode, rooted in endogenous mechanisms of the individual.

The Metastability of the Double-Tripod Gait in Locust Locomotion.

Insect locomotion represents a fundamental example of neuronal oscillating circuits generating different motor patterns or gaits by controlling their phase coordination. Walking gaits are assumed to represent stable states of the system, often modeled as coupled oscillators. This view is challenged, however, by recent experimental observations, in which in vitro locust preparations consistently converged to synchronous rhythms (all legs oscillating as one), a locomotive pattern never seen in vivo. To reconcile this inconsistency, we developed a modeling framework to capture the trade-off between the two competing mechanisms: the endogenous neuronal circuitry, expressed in vitro, and the feedback mechanisms from sensory and descending inputs, active only in vivo. We show that the ubiquitously observed double-tripod walking gait emerges precisely from this balance. The outcome is a short-lived meta-stable double-tripod gait, which transitions and alternates with stable idling, thus recovering the observed intermittent bouts of locomotion, typical of many insects' locomotion behavior.

The functional connectivity between the locust leg pattern generators and the subesophageal ganglion higher motor center.

Higher motor centers and central pattern generators (CPGs) interact in the control of coordinated leg movements during locomotion throughout the animal kingdom. The subesophageal ganglion (SEG) is one of the insect head ganglia reported to have a role in the control of walking behavior. Here we explored the functional relations between the SEG and the thoracic leg CPGs in the desert locust. Backfill staining revealed about 300 SEG descending interneurons (DINs) altogether. Recordings from an in-vitro isolated chain of thoracic ganglia, with intact or severed connections to the SEG, during pharmacological activation were used to determine how the SEG affects the centrally generated motor output to the legs. The SEG was demonstrated to both activate leg CPGs and synchronize their bilateral activity. The role of the SEG in insect locomotion is discussed in light of these findings.

Ex vivo recordings reveal desert locust forelimb control is asymmetric.

Lateralized behaviours are widespread among the animals, including insects with their miniature brains, perhaps being a way of maximising neural capacity (reviewed in [1,2]). However, evidence for functional asymmetries in the neural circuitry itself is scarce. Here, using bilateral simultaneous recordings from the ex vivo nervous system of desert locusts, we show that the neural control of their forelimbs is asymmetric. This asymmetry was retained throughout the experimental period and either with or without the suboesophageal ganglion (SOG). These findings provide evidence for hard-wired neural sidedness and contribute to our understanding of the lateralization observed in in-vivo motor behaviours.

The use of MEMRI for monitoring central nervous system activity during intact insect walking.

BACKGROUND: Monitoring neuronal activity in the intact behaving animal is most desired in neuroethological research, yet it is rarely straightforward or even feasible. Here we present the use of manganese enhanced magnetic resonance imaging (MEMRI), a technique allowing monitoring the activity of an animal's nervous system during specific behavioral patterns. Using MEMRI we were able to show activity in different ganglia of the central nervous system of intact locusts during walking. RESULTS: We injected two groups of locusts with manganese, which serves as a magnetic contrast agent. One group was forced to walk on a treadmill for two hours, while the other was immobilized and served as a control. Subsequently, all animals were scanned in a T1 MRI protocol, and the accumulation of manganese in the neuronal tissues that were active during walking was demonstrated by comparing the scans of the two groups. Two neuronal sites showed significantly higher T1 signal in the walking locusts compared to the immobilized ones: the prothoracic ganglion, which locally controls the front legs, and the subesophageal ganglion, a head ganglion which takes part in initiation and maintenance of walking. CONCLUSION: MEMRI is a potent, non-invasive technique for monitoring neuronal activity in intact locusts, and arthropods in general. Specifically, it provides a promising way for revealing the role of central and high-order neuronal structures in motor behaviors such as walking.

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.

Rigidity and Flexibility: The Central Basis of Inter-Leg Coordination in the Locust.

Many motor behaviors, and specifically locomotion, are the product of an intricate interplay between neuronal oscillators known as central pattern generators (CPGs), descending central commands, and sensory feedback loops. The relative contribution of each of these components to the final behavior determines the trade-off between fixed movements and those that are carefully adapted to the environment. Here we sought to decipher the endogenous, default, motor output of the CPG network controlling the locust legs, in the absence of any sensory or descending influences. We induced rhythmic activity in the leg CPGs in isolated nervous system preparations, using different application procedures of the muscarinic agonist pilocarpine. We found that the three thoracic ganglia, each controlling a pair of legs, have different inherent bilateral coupling. Furthermore, we found that the pharmacological activation of one ganglion is sufficient to induce activity in the other, untreated, ganglia. Each ganglion was thus capable to impart its own bilateral inherent pattern onto the other ganglia via a tight synchrony among the ipsilateral CPGs. By cutting a connective and severing the lateral-longitudinal connections, we were able to uncouple the oscillators' activity. While the bilateral connections demonstrated a high modularity, the ipsilateral CPGs maintained a strict synchronized activity. These findings suggest that the central infrastructure behind locust walking features both rigid elements, which presumably support the generation of stereotypic orchestrated leg movements, and flexible elements, which might provide the central basis for adaptations to the environment and to higher motor commands.

The effect of octopamine on the locust stomatogastric nervous system.

Octopamine (OA) is a prominent neuromodulator of invertebrate nervous systems, influencing multiple physiological processes. Among its many roles in insects are the initiation and maintenance of various rhythmic behaviors. Here, the neuromodulatory effects of OA on the components of the locust stomatogastric nervous system were studied, and one putative source of OA modulation of the system was identified. Bath application of OA was found to abolish the endogenous rhythmic output of the fully isolated frontal ganglion (FG), while stimulating motor activity of the fully isolated hypocerebral ganglion (HG). OA also induced rhythmic movements in a foregut preparation with intact HG innervation. Complex dose-dependent effects of OA on interconnected FG-HG preparations were seen: 10(-5) M OA accelerated the rhythmic activity of both the HG and FG in a synchronized manner, while 10(-4) M OA decreased both rhythms. Intracellular stimulation of an identified octopaminergic dorsal unpaired median neuron in the subesophageal ganglion was found to exert a similar effect on the FG motor output as that of OA application. Our findings suggest a mechanism of regulation of insect gut patterns and feeding-related behavior during stress and times of high energy demand.