The Role of Central Complex Neurons in Prey Detection and Tracking in the Freely Moving Praying Mantis (Tenodera sinensis).

Complex tasks like hunting moving prey in an unpredictable environment require high levels of motor and sensory integration. An animal needs to detect and track suitable prey objects, measure their distance and orientation relative to its own position, and finally produce the correct motor output to approach and capture the prey. In the insect brain, the central complex (CX) is one target area where integration is likely to take place. In this study, we performed extracellular multi-unit recordings on the CX of freely hunting praying mantises (Tenodera sinensis). Initially, we recorded the neural activity of freely moving mantises as they hunted live prey. The recordings showed activity in cells that either reflected the mantis's own movements or the actions of a prey individual, which the mantises focused on. In the latter case, the activity increased as the prey moved and decreased when it stopped. Interestingly, cells ignored the movement of the other prey than the one to which the mantis attended. To obtain quantitative data, we generated simulated prey targets presented on an LCD screen positioned below the clear floor of the arena. The simulated target oscillated back and forth at various angles and distances. We identified populations of cells whose activity patterns were strongly linked to the appearance, movement, and relative position of the virtual prey. We refer to these as sensory responses. We also found cells whose activity preceded orientation movement toward the prey. We call these motor responses. Some cells showed both sensory and motor properties. Stimulation through tetrodes in some of the preparations could also generate similar movements. These results suggest the crucial importance of the CX to prey-capture behavior in predatory insects like the praying mantis and, hence, further emphasize its role in behaviorally and ecologically relevant contexts.

Context-dependent effects on attack and defense behaviors in the praying mantis Tenodera sinensis.

Most behavior needs to strike a balance between the competing needs to find food and protect an animal from predators. The factors that influence this balance and the resulting behavior are not well understood in many animals. Here, we examined these influences in the praying mantis Tenodera sinensis by presenting perching individuals with alternating sinusoidally moving prey-like stimuli and rapidly expanding looming stimuli then scoring their behavior on a defensive-aggressive scale. In this way, we tested the hypothesis that such behaviors are highly context dependent. Specifically, we found that defensive responses, which are normally very consistent, are decreased in magnitude if the animal has just performed an aggressive response to the previous sinusoid. A thrash behavior not normally seen with looming alone was often seen following aggression. In thrashing, the animal tries to push the looming stimulus away. Thrashing almost exclusively followed aggressive responses to the sinusoid stimulus. Moreover, aggression levels were found to shift from low to high and back to low as adult animals aged and, in general, female mantises were more aggressive than males. Finally, the specific nature of the mid-life spike in aggressive behaviors differed according to whether the animals were lab raised or caught in the wild. Lab-raised animals showed roughly equal amounts of increased attention to the stimulus and very aggressive strike behaviors, whereas wild-caught animals tended to either ignore the stimulus or react very aggressively with strikes. Therefore, our hypothesis regarding context-dependent effects was supported, with all four factors influencing the behaviors that were studied.

Insect neuroethology: Flight constructs a compass for monarch migration.

A new study shows that a brain map in the monarch butterfly can be re-drawn during flight. Migrating butterflies integrate efferent and visual signals to create an adaptable compass in their central brain.

Internal state effects on behavioral shifts in freely behaving praying mantises (Tenodera sinensis).

How we interact with our environment largely depends on both the external cues presented by our surroundings and the internal state from within. Internal states are the ever-changing physiological conditions that communicate the immediate survival needs and motivate the animal to behaviorally fulfill them. Satiety level constitutes such a state, and therefore has a dynamic influence on the output behaviors of an animal. In predatory insects like the praying mantis, hunting tactics, grooming, and mating have been shown to change hierarchical organization of behaviors depending on satiety. Here, we analyze behavior sequences of freely hunting praying mantises (Tenodera sinensis) to explore potential differences in sequential patterning of behavior as a correlate of satiety. First, our data supports previous work that showed starved praying mantises were not just more often attentive to prey, but also more often attentive to further prey. This was indicated by the increased time fraction spent in attentive bouts such as prey monitoring, head turns (to track prey), translations (closing the distance to the prey), and more strike attempts. With increasing satiety, praying mantises showed reduced time in these behaviors and exhibited them primarily towards close-proximity prey. Furthermore, our data demonstrates that during states of starvation, the praying mantis exhibits a stereotyped pattern of behavior that is highly motivated by prey capture. As satiety increased, the sequenced behaviors became more variable, indicating a shift away from the necessity of prey capture to more fluid presentations of behavior assembly.

Predatory behavior changes with satiety or increased insulin levels in the praying mantis (Tenodera sinensis).

At any given moment, behavior is controlled by a combination of external stimuli and an animal's internal state. As physiological conditions change, vastly different behaviors might result from the same stimuli. For example, the motivation to hunt and hunting strategy are influenced by satiety. Here, we describe how sensory responsiveness and motor activity of a praying mantis (Tenodera sinensis) change as the insect feeds, leading to an altered hunting strategy. We further show that these changes can be induced by injection of insulin, which likely functions as a metabotropic indicator. Praying mantises directed their attention toward real and simulated prey less often as they fed and became sated. The range of distance and azimuth at which prey was detected decreased as did pursuit of prey, while opportunistic close-range attacks persisted. Together, these sensorimotor changes are indicative of a behavioral paradigm shift from 'pursuit' to 'ambush'. A similar effect was induced in starved praying mantises injected with 0.05 ml of 200 µg ml-1 bovine insulin. These experiments showed that insulin injection into the circulating hemolymph is sufficient to decrease prey orientation as well as in prey-directed locomotor behaviors (tracking and pursuit). The effects of prey consumption and insulin injection were similarly dose dependent. These results suggest that insulin is a signal of internal, physiological conditions that can modify responses to external stimuli. A change in hunting strategy thus results from coordinated effects of a neurohormone on a set of independent sensorimotor processes and the overall activity level of the animal.

Mantisbot is a robotic model of visually guided motion in the praying mantis.

Insects use highly distributed nervous systems to process exteroception from head sensors, compare that information with state-based goals, and direct posture or locomotion toward those goals. To study how descending commands from brain centers produce coordinated, goal-directed motion in distributed nervous systems, we have constructed a conductance-based neural system for our robot MantisBot, a 29 degree-of-freedom, 13.3:1 scale praying mantis robot. Using the literature on mantis prey tracking and insect locomotion, we designed a hierarchical, distributed neural controller that establishes the goal, coordinates different joints, and executes prey-tracking motion. In our controller, brain networks perceive the location of prey and predict its future location, store this location in memory, and formulate descending commands for ballistic saccades like those seen in the animal. The descending commands are simple, indicating only 1) whether the robot should walk or stand still, and 2) the intended direction of motion. Each joint's controller uses the descending commands differently to alter sensory-motor interactions, changing the sensory pathways that coordinate the joints' central pattern generators into one cohesive motion. Experiments with one leg of MantisBot show that visual input produces simple descending commands that alter walking kinematics, change the walking direction in a predictable manner, enact reflex reversals when necessary, and can control both static posture and locomotion with the same network.

Spatial Navigation and the Central Complex: Sensory Acquisition, Orientation, and Motor Control.

Cockroaches are scavengers that forage through dark, maze-like environments. Like other foraging animals, for instance rats, they must continually asses their situation to keep track of targets and negotiate barriers. While navigating a complex environment, all animals need to integrate sensory information in order to produce appropriate motor commands. The integrated sensory cues can be used to provide the animal with an environmental and contextual reference frame for the behavior. To successfully reach a goal location, navigational cues continuously derived from sensory inputs have to be utilized in the spatial guidance of motor commands. The sensory processes, contextual and spatial mechanisms, and motor outputs contributing to navigation have been heavily studied in rats. In contrast, many insect studies focused on the sensory and/or motor components of navigation, and our knowledge of the abstract representation of environmental context and spatial information in the insect brain is relatively limited. Recent reports from several laboratories have explored the role of the central complex (CX), a sensorimotor region of the insect brain, in navigational processes by recording the activity of CX neurons in freely-moving insects and in more constrained, experimenter-controlled situations. The results of these studies indicate that the CX participates in processing the temporal and spatial components of sensory cues, and utilizes these cues in creating an internal representation of orientation and context, while also directing motor control. Although these studies led to a better understanding of the CX's role in insect navigation, there are still major voids in the literature regarding the underlying mechanisms and brain regions involved in spatial navigation. The main goal of this review is to place the above listed findings in the wider context of animal navigation by providing an overview of the neural mechanisms of navigation in rats and summarizing and comparing our current knowledge on the CX's role in insect navigation to these processes. By doing so, we aimed to highlight some of the missing puzzle pieces in insect navigation and provide a different perspective for future directions.

Cellular Basis of Head Direction and Contextual Cues in the Insect Brain.

Animals rely upon integrated sensory information for spatial navigation. A question of wide importance in navigation is how sensory cues get transformed into neural codes that represent the animal's orientation within its proximal environment. Here, we investigated the possibility of head-direction coding in the central complex of the cockroach, Blaberus discoidalis. We used extracellular recordings in restrained animals that were rotated on a platform relative to a fixed landmark. The passive rotations allowed us to test for head-direction coding in the absence of self-generated motion cues. Our results indicate that individual cells in the central complex encode the animal's heading relative to a landmark's position in several ways. In some cells, directional tuning was established even in the absence of visual cues, suggesting that the directional code can be maintained solely based on the internal motion cues derived from the passive rotations. Additionally, some cells in the central complex encoded rotation-direction history, a navigational context cue, by increasing or decreasing the firing rate during the stationary periods following clockwise or counterclockwise rotations. Together, these results unveil head-direction cell-like activity in the insect central complex, which highly resemble similarly functioning cells in the mammalian brain that encode head direction. We predict that the observed head-orientation coding and directionally sensitive cells are essential components of the brain circuitry mediating insect navigation.

Neuromechanical model of praying mantis explores the role of descending commands in pre-strike pivots.

Praying mantises hunt by standing on their meso- and metathoracic legs and using them to rotate and translate (together, 'pivot') their bodies toward prey. We have developed a neuromechanical software model of the praying mantis Tenodera sinensis to use as a platform for testing postural controllers that the animal may use while hunting. Previous results showed that a feedforward model was insufficient for capturing the diversity of posture observed in the animal (Szczecinski et al 2014 Biomimetic and Biohybrid Syst. 3 296-307). Therefore we have expanded upon this model to make a flexible controller with feedback that more closely mimics the animal. The controller actuates 24 joints in the legs of a dynamical model to orient the head and translate the thorax toward prey. It is controlled by a simulation of nonspiking neurons assembled as a highly simplified version of networks that may exist in the mantid central complex and thoracic ganglia. Because of the distributed nature of these networks, we hypothesize that descending commands that orient the mantis toward prey may be simple direction-of-intent signals, which are turned into motor commands by the structure of low-level networks in the thoracic ganglia. We verify this through a series of experiments with the model. It captures the speed and range of mantid pivots as reported in other work (Yamawaki et al 2011 J. Insect Physiol. 57 1010-6). It is capable of pivoting toward prey from a variety of initial postures, as seen in the animal. Finally, we compare the model's joint kinematics during pivots to preliminary 3D kinematics collected from Tenodera.

Central-complex control of movement in the freely walking cockroach.

To navigate in the world, an animal's brain must produce commands to move, change direction, and negotiate obstacles. In the insect brain, the central complex integrates multiple forms of sensory information and guides locomotion during behaviors such as foraging, climbing over barriers, and navigating to memorized locations. These roles suggest that the central complex influences motor commands, directing the appropriate movement within the current context. Such commands are ultimately carried out by the limbs and must therefore interact with pattern generators and reflex circuits that coordinate them. Recent studies have described how neurons of the central complex encode sensory information: neurons subdivide the space around the animal, encoding the direction or orientation of stimuli used in navigation. Does a similar central-complex code directing movement exist, and if so, how does it effect changes in the control of limbs? Recording from central-complex neurons in freely walking cockroaches (Blaberus discoidalis), we identified classes of movement-predictive cells selective for slow or fast forward walking, left or right turns, or combinations of forward and turning speeds. Stimulation through recording wires produced consistent trajectories of forward walking or turning in these animals, and those that elicited turns also altered an inter-joint reflex to a pattern resembling spontaneous turning. When an animal transitioned to climbing over an obstacle, the encoding of movement in this new context changed for a subset of cells. These results indicate that encoding of movement in the central complex participates in motor control by a distributed, flexible code targeting limb reflex circuits.

How cockroaches exploit tactile boundaries to find new shelters.

Animals such as cockroaches depend on exploration of unknown environments, and their strategies may inspire robotic approaches. We have previously shown that cockroach behavior, with respect to shelters and the walls of an otherwise empty arena, can be captured with a stochastic state-based algorithm. We call this algorithm RAMBLER, randomized algorithm mimicking biased lone exploration in roaches. In this work, we verified and extended this model by adding a barrier in the previously used arena and conducted more cockroach experiments. In two arena configurations, our simulated model's path length distribution was similar to the experimental distribution (mean experimental path length 3.4 and 3.2 m, mean simulated path length 3.9 and 3.3 m). By analyzing cockroach behavior before, along, and at the end of the barrier, we have generalized RAMBLER to address arbitrarily complex 2D mazes. For biology, this is an abstract behavioral model of a decision-making process in the cockroach brain. For robotics, this is a strategy that may improve exploration for goals, especially in unpredictable environments with non-convex obstacles. Generally, cockroach behavior seems to recommend variability in the absence of planning, and following paths defined by walls.

Encoding wide-field motion and direction in the central complex of the cockroach Blaberus discoidalis.

In the arthropod brain, the central complex (CX) receives various forms of sensory signals and is associated with motor functions, but its precise role in behavior is controversial. The optomotor response is a highly conserved turning behavior directed by visual motion. In tethered cockroaches, 20% procaine injected into the CX reversibly blocked this behavior. We then used multichannel extracellular recording to sample unit activity in the CX in response to wide-field visual motion stimuli, moving either horizontally or vertically at various temporal frequencies. For the 401 units we sampled, we identified five stereotyped response patterns: tonically inhibited or excited responses during motion, phasically inhibited or excited responses at the initiation of motion, and phasically excited responses at the termination of motion. Sixty-seven percent of the units responded to horizontal motion, while only 19% responded to vertical motion. Thirty-eight percent of responding units were directionally selective to horizontal motion. Response type and directional selectivity were sometimes conditional with other stimulus parameters, such as temporal frequency. For instance, 16% of the units that responded tonically to low temporal frequencies responded phasically to high temporal frequencies. In addition, we found that 26% of wide-field motion responding units showed a periodic response that was entrained to the temporal frequency of the stimulus. Our results show a diverse population of neurons within the CX that are variably tuned to wide-field motion parameters. Our behavioral data further suggest that such CX activity is required for effective optomotor responses.

Extracellular wire tetrode recording in brain of freely walking insects.

Increasing interest in the role of brain activity in insect motor control requires that we be able to monitor neural activity while insects perform natural behavior. We previously developed a technique for implanting tetrode wires into the central complex of cockroach brains that allowed us to record activity from multiple neurons simultaneously while a tethered cockroach turned or altered walking speed. While a major advance, tethered preparations provide access to limited behaviors and often lack feedback processes that occur in freely moving animals. We now present a modified version of that technique that allows us to record from the central complex of freely moving cockroaches as they walk in an arena and deal with barriers by turning, climbing or tunneling. Coupled with high speed video and cluster cutting, we can now relate brain activity to various parameters of the movement of freely behaving insects.

A neuromechanical simulation of insect walking and transition to turning of the cockroach Blaberus discoidalis.

A neuromechanical simulation of the cockroach Blaberus discoidalis was developed to explore changes in locomotion when the animal transitions from walking straight to turning. The simulation was based upon the biological data taken from three sources. Neural circuitry was adapted from the extensive literature primarily obtained from the studies of neural connections within thoracic ganglia of stick insect and adapted to cockroach. The 3D joint kinematic data on straight, forward walking for cockroach were taken from a paper that describes these movements in all joints simultaneously as the cockroach walked on an oiled-plate tether (Bender et al. in PloS one 5(10):1-15, 2010b). Joint kinematics for turning were only available for some leg joints (Mu and Ritzmann in J Comp Physiol A Neuroethol Sens Neural Behav Physiol 191(11):1037-54, 2005) and thus had to be obtained using the methods that were applied for straight walking by Bender et al. (PloS one 5(10):1-15, 2010b). Once walking, inside turning, and outside turning were characterized, phase and amplitude changes for each joint of each leg were quantified. Apparent reflex reversals and joint activity changes were used to modify sensory coupling pathways between the CPG at each joint of the simulation. Oiled-plate experiments in simulation produced tarsus trajectories in stance similar to those seen in the animal. Simulations including forces that would be experienced if the insect was walking freely (i.e., weight support and friction) again produced similar results. These data were not considered during the design of the simulation, suggesting that the simulation captures some key underlying the principles of walking, turning, and transitioning in the cockroach. In addition, since the nervous system was modeled with realistic neuron models, biologically plausible reflex reversals are simulated, motivating future neurobiological research.

Neural activity in the central complex of the cockroach brain is linked to turning behaviors.

An animal moving through complex terrain must consider sensory cues around it and alter its movements accordingly. In the arthropod brain, the central complex (CC) receives highly preprocessed sensory information and sends outputs to premotor regions, suggesting that it may play a role in the central control of oriented locomotion. We performed tetrode recordings within the CC in cockroaches walking on an air-suspended ball to examine the role of the CC in turning behaviors. When a rod was placed near the cockroach's head, the cockroach touched the rod repeatedly with one or both antennae before locomotion was initiated. Some CC units responded to self-generated antennal contact with the object, but at lower levels compared with externally imposed antennal stimulation. The neural activity of other CC units responded to locomotion. We found that some CC units showed discrete firing fields corresponding to specific locomotion states. We also found that changes in firing rate of some CC units preceded changes in turning speed in one direction but not the other. Furthermore, such biased units were located in the side of the brain ipsilateral to the direction of the turning speed they could predict. Moreover, electrical stimulation of the CC elicited or modified locomotion, and the direction of some evoked locomotion could be predicted by the response property of locomotion-predictive units near the stimulation site. Therefore, our results suggest that, at the population level, asymmetrical activity in the CC precedes and influences turning behavior.

Deciding which way to go: how do insects alter movements to negotiate barriers?

Animals must routinely deal with barriers as they move through their natural environment. These challenges require directed changes in leg movements and posture performed in the context of ever changing internal and external conditions. In particular, cockroaches use a combination of tactile and visual information to evaluate objects in their path in order to effectively guide their movements in complex terrain. When encountering a large block, the insect uses its antennae to evaluate the object's height then rears upward accordingly before climbing. A shelf presents a choice between climbing and tunneling that depends on how the antennae strike the shelf; tapping from above yields climbing, while tapping from below causes tunneling. However, ambient light conditions detected by the ocelli can bias that decision. Similarly, in a T-maze turning is determined by antennal contact but influenced by visual cues. These multi-sensory behaviors led us to look at the central complex as a center for sensori-motor integration within the insect brain. Visual and antennal tactile cues are processed within the central complex and, in tethered preparations, several central complex units changed firing rates in tandem with or prior to altered step frequency or turning, while stimulation through the implanted electrodes evoked these same behavioral changes. To further test for a central complex role in these decisions, we examined behavioral effects of brain lesions. Electrolytic lesions in restricted regions of the central complex generated site specific behavioral deficits. Similar changes were also found in reversible effects of procaine injections in the brain. Finally, we are examining these kinds of decisions made in a large arena that more closely matches the conditions under which cockroaches forage. Overall, our studies suggest that CC circuits may indeed influence the descending commands associated with navigational decisions, thereby making them more context dependent.

An implantable biofuel cell for a live insect.

A biofuel cell incorporating a bienzymatic trehalase|glucose oxidase trehalose anode and a bilirubin oxidase dioxygen cathode using Os complexes grafted to a polymeric backbone as electron relays was designed and constructed. The specific power densities of the biofuel cell implanted in a female Blaberus discoidalis through incisions into its abdomen yielded maximum values of ca. 55 µW/cm(2) at 0.2 V that decreased by only ca. 5% after ca. 2.5 h of operation.

Kinematic and behavioral evidence for a distinction between trotting and ambling gaits in the cockroach Blaberus discoidalis.

Earlier observations had suggested that cockroaches might show multiple patterns of leg coordination, or gaits, but these were not followed by detailed behavioral or kinematic measurements that would allow a definite conclusion. We measured the walking speeds of cockroaches exploring a large arena and found that the body movements tended to cluster at one of two preferred speeds, either very slow (<10 cm s(-1)) or fairly fast (∼30 cm s(-1)). To highlight the neural control of walking leg movements, we experimentally reduced the mechanical coupling among the various legs by tethering the animals and allowing them to walk in place on a lightly oiled glass plate. Under these conditions, the rate of stepping was bimodal, clustering at fast and slow speeds. We next used high-speed videos to extract three-dimensional limb and joint kinematics for each segment of all six legs. The angular excursions and three-dimensional motions of the leg joints over the course of a stride were variable, but had different distributions in each gait. The change in gait occurs at a Froude number of ∼0.4, a speed scale at which a wide variety of animals show a transition between walking and trotting. We conclude that cockroaches do have multiple gaits, with corresponding implications for the collection and interpretation of data on the neural control of locomotion.

Computer-assisted 3D kinematic analysis of all leg joints in walking insects.

High-speed video can provide fine-scaled analysis of animal behavior. However, extracting behavioral data from video sequences is a time-consuming, tedious, subjective task. These issues are exacerbated where accurate behavioral descriptions require analysis of multiple points in three dimensions. We describe a new computer program written to assist a user in simultaneously extracting three-dimensional kinematics of multiple points on each of an insect's six legs. Digital video of a walking cockroach was collected in grayscale at 500 fps from two synchronized, calibrated cameras. We improved the legs' visibility by painting white dots on the joints, similar to techniques used for digitizing human motion. Compared to manual digitization of 26 points on the legs over a single, 8-second bout of walking (or 106,496 individual 3D points), our software achieved approximately 90% of the accuracy with 10% of the labor. Our experimental design reduced the complexity of the tracking problem by tethering the insect and allowing it to walk in place on a lightly oiled glass surface, but in principle, the algorithms implemented are extensible to free walking. Our software is free and open-source, written in the free language Python and including a graphical user interface for configuration and control. We encourage collaborative enhancements to make this tool both better and widely utilized.

Neural activity in the central complex of the insect brain is linked to locomotor changes.

Animals negotiating complex natural terrain must consider cues around them and alter movement parameters accordingly. In the arthropod brain, the central complex (CC) receives bilateral sensory relays and sits immediately upstream of premotor areas, suggesting that it may be involved in the context-dependent control of behavior. In previous studies, CC neurons in various insects responded to visual, chemical, and mechanical stimuli, and genetic or physical lesions affected locomotor behaviors. Additionally, electrical stimulation of the CC led to malformed chirping movements by crickets, and pharmacological stimulation evoked stridulation in grasshoppers, but no more precise relationship has been documented between neural activity in the CC and movements in a behaving animal. We performed tetrode recordings from the CC of cockroaches walking in place on a slippery surface. Neural activity in the CC was strongly correlated with, and in some cases predictive of, stepping frequency. Electrical stimulation of these areas also evoked or modified walking. Many of the same neural units responded to tactile antennal stimulation while the animal was standing still but became unresponsive during walking. Therefore, these CC units are unlikely to be reporting only sensory signals, but their activity may be directing changes in locomotion based on sensory inputs.