Cercal System Research Articles

Roles of neural communication between the brain and thoracic ganglia in the selection and regulation of the cricket escape behavior

To survive a predator’s attack, prey animals must exhibit escape responses that are appropriately regulated in terms of their moving speed, distance, and direction. Insect locomotion is considered to be controlled by an interaction between the brain, which is involved in behavioral decision-making, and the thoracic ganglia (TG), which are primary motor centers. However, it remains unknown which descending and ascending signals between these neural centers are involved in the regulation of the escape behavior. We addressed the distinct roles of the brain and TG in the wind-elicited escape behavior of crickets by assessing the effects of partial ablation of the intersegmental communications on escape responses. We unilaterally cut the ventral nerve cord (VNC) at different locations, between the brain and TG, or between the TG and terminal abdominal ganglion (TAG), a primary sensory center of the cercal system. The partial ablation of ascending signals to the brain greatly reduced the jumping response rather than running, indicating that sensory information processing in the brain is essential for the choice of escape responses. The ablation of descending signals from the brain to the TG impaired locomotor performance and directional control of the escape responses, suggesting that locomotion in the escape behavior largely depends on the descending signals from the brain. Finally, the extracellular recording from the cervical VNC indicated a difference in the descending activities preceding the escape responses between running and jumping. Our results demonstrated that the brain sends the descending signals encoding the behavioral choice and locomotor regulation to the TG, while the TG seem to have other specific roles, such as in the preparation of escape movement.

Transcriptional expression changes during compensatory plasticity in the\xa0terminal ganglion of the adult cricket Gryllus bimaculatus

BackgroundDamage to the adult central nervous system often leads to long-term disruptions in function due to the limited capacity for neurological recovery. The central nervous system of the Mediterranean field cricket, Gryllus bimaculatus, shows an unusual capacity for compensatory plasticity, most obviously in the auditory system and the cercal escape system. In both systems, unilateral sensory disruption leads the central circuitry to compensate by forming and/or strengthening connections with the contralateral sensory organ. While this compensatory plasticity in the auditory system relies on robust dendritic sprouting and novel synapse formation, the compensatory plasticity in the cercal escape circuitry shows little obvious dendritic sprouting and instead may rely on shifts in excitatory and inhibitory synaptic strength.ResultsIn order to better understand what types of molecular pathways might underlie this compensatory shift in the cercal system, we used a multiple k-mer approach to assemble a terminal ganglion transcriptome that included ganglia collected one, three, and 7 days after unilateral cercal ablation in adult, male animals. We performed differential expression analysis using EdgeR and DESeq2 and examined Gene Ontologies to identify candidates potentially involved in this plasticity. Enriched GO terms included those related to the ubiquitin-proteosome protein degradation system, chromatin-mediated transcriptional pathways, and the GTPase-related signaling system.ConclusionFurther exploration of these GO terms will provide a clearer picture of the processes involved in compensatory recovery of the cercal escape system in the cricket and can be compared and contrasted with the distinct pathways that have been identified upon deafferentation of the auditory system in this same animal.

Post-molting development of wind-elicited escape behavior in the cricket

Arthropods including insects grow through several developmental stages by molting. The abrupt changes in their body size and morphology accompanying the molting are responsible for the developmental changes in behavior. While in holometabolous insects, larval behaviors are transformed into adult-specific behaviors with drastic changes in nervous system during the pupal stage, hemimetabolous insects preserve most innate behaviors whole life long, which allow us to trace the maturation process of preserved behaviors after the changes in body. Wind-elicited escape behavior is one of these behaviors and mediated by cercal system, which is a mechanosensory organ equipped by all stages of nymph in orthopteran insects like crickets. However, the maturation process of the escape behavior after the molt is unclear. In this study, we examined time-series of changes in the wind-elicited escape behavior just after the imaginal molt in the cricket. The locomotor activities are developed over the elapsed time, and matured 24h after the molt. In contrast, a stimulus-angle dependency of moving direction was unchanged over time, meaning that the cercal sensory system detecting airflow direction was workable immediately after the molt, independent from the behavioral maturation. The post-molting development of the wind-elicited behavior was considered to result not simply from maturation of the exoskeleton or musculature because the escape response to heat-shock stimulus did not change after the molt. No effect of a temporal immobilization after the imaginal molt on the maturation of the wind-elicited behavior also implies that the maturation may be innately programmed without experience of locomotion.

Impact of cercal air currents on singing motor pattern generation in the cricket (Gryllus bimaculatus DeGeer).

The cercal system of crickets detects low-frequency air currents produced by approaching predators and self-generated air currents during singing, which may provide sensory feedback to the singing motor network. We analyzed the effect of cercal stimulation on singing motor pattern generation to reveal the response of a singing interneuron to predator-like signals and to elucidate the possible role of self-generated air currents during singing. In fictive singing males, we recorded an interneuron of the singing network while applying air currents to the cerci; additionally, we analyzed the effect of abolishing the cercal system in freely singing males. In fictively singing crickets, the effect of short air stimuli is either to terminate prematurely or to lengthen the interchirp interval, depending on their phase in the chirp cycle. Within our stimulation paradigm, air stimuli of different velocities and durations always elicited an inhibitory postsynaptic potential in the singing interneuron. Current injection in the singing interneuron elicited singing motor activity, even during the air current-evoked inhibitory input from the cercal pathway. The disruptive effects of air stimuli on the fictive singing pattern and the inhibitory response of the singing interneuron point toward the cercal system being involved in initiating avoidance responses in singing crickets, according to the established role of cerci in a predator escape pathway. After abolishing the activity of the cercal system, the timing of natural singing activity was not significantly altered. Our study provides no evidence that self-generated cercal sensory activity has a feedback function for singing motor pattern generation.

Corollary discharge inhibition of wind-sensitive cercal giant interneurons in the singing field cricket.

Crickets carry wind-sensitive mechanoreceptors on their cerci, which, in response to the airflow produced by approaching predators, triggers escape reactions via ascending giant interneurons (GIs). Males also activate their cercal system by air currents generated due to the wing movements underlying sound production. Singing males still respond to external wind stimulation, but are not startled by the self-generated airflow. To investigate how the nervous system discriminates sensory responses to self-generated and external airflow, we intracellularly recorded wind-sensitive afferents and ventral GIs of the cercal escape pathway in fictively singing crickets, a situation lacking any self-stimulation. GI spiking was reduced whenever cercal wind stimulation coincided with singing motor activity. The axonal terminals of cercal afferents showed no indication of presynaptic inhibition during singing. In two ventral GIs, however, a corollary discharge inhibition occurred strictly in phase with the singing motor pattern. Paired intracellular recordings revealed that this inhibition was not mediated by the activity of the previously identified corollary discharge interneuron (CDI) that rhythmically inhibits the auditory pathway during singing. Cercal wind stimulation, however, reduced the spike activity of this CDI by postsynaptic inhibition. Our study reveals how precisely timed corollary discharge inhibition of ventral GIs can prevent self-generated airflow from triggering inadvertent escape responses in singing crickets. The results indicate that the responsiveness of the auditory and wind-sensitive pathway is modulated by distinct CDIs in singing crickets and that the corollary discharge inhibition in the auditory pathway can be attenuated by cercal wind stimulation.

Neural responses from the filiform receptor neuron afferents of the wind-sensitive cercal system in three cockroach species

The wind-sensitive insect cercal system is involved in many important behaviors, such as initiating terrestrial escape responses and providing sensory feedback during flight. The occurrence of these behaviors vary in cockroach species Periplaneta americana (strong terrestrial response and flight), Blaberus craniifer (weak terrestrial response and flight), and Gromphodorhina portentosa (no terrestrial response and no flight). A previous study focusing on wind-sensitive interneuron (WSI) responses demonstrated that variations in sensory processing of wind information accompany these behavioral differences. In this study, we recorded extracellularly from the cercal nerve to characterize filiform afferent population responses to different wind velocities to investigate how sensory processing differs across these species at the initial encoding of wind. We compared these results and responses from the WSI population to examine information transfer at the first synapse. Our main results were: (1) G. portentosa had the weakest responses of the three species over the stimulus duration and possessed the smallest cerci with the least filiform hair receptors of the three species; (2) B. craniifer filiform responses were similar to or greater than P. americana responses even though B. craniifer possessed smaller cerci with less filiform hair receptors than P. americana; (3) the greater filiform afferent responses in B. craniifer, including a larger amplitude second positive peak compared to the other two species, suggest more synchronous activity between filiform afferents in this species; (4) the transfer of information at the first synapse appears to be similar in both P. americana and G. portentosa, but different in B. craniifer.

Neural responses from the wind-sensitive interneuron population in four cockroach species

The wind-sensitive insect cercal sensory system is involved in important behaviors including predator detection and initiating terrestrial escape responses as well as flight maintenance. However, not all insects possessing a cercal system exhibit these behaviors. In cockroaches, wind evokes strong terrestrial escape responses in Periplaneta americana and Blattella germanica, but only weak escape responses in Blaberus craniifer and no escape responses in Gromphadorhina portentosa. Both P. americana and B. craniifer possesses pink flight muscles correlated with flight ability while B. germanica possesses white flight muscles that cannot support flight and G. portentosa lacks wings. These different behavioral combinations could correlate with differences in sensory processing of wind information by the cercal system. In this study, we focused on the wind-sensitive interneurons (WSIs) since they provide input to the premotor/motor neurons that influence terrestrial escape and flight behavior. Using extracellular recordings, we characterized the responses from the WSI population by generating stimulus–response (S–R) curves and examining spike firing rates. Using cluster analysis, we also examined the activity of individual units (four per species, though not necessarily homologous) comprising the population response in each species. Our main results were: (1) all four species possessed ascending WSIs in the abdominal connectives; (2) wind elicited the weakest WSI responses (lowest spike counts and spike rates) in G. portentosa; (3) wind elicited WSI responses in B. craniifer that were greater than P. americana or B. germanica; (4) the activity of four individual units comprising the WSI population response in each species was similar across species.

Responses of cricket cercal interneurons to realistic naturalistic stimuli in the field

The ability of the insect cercal system to detect approaching predators has been studied extensively in the laboratory and in the field. Some previous studies have assessed the extent to which sensory noise affects the operational characteristics of the cercal system, but these studies have only been carried out in laboratory settings using white noise stimuli of unrealistic nature. Using a piston mimicking the natural airflow of an approaching predator, we recorded the neural activity through the abdominal connectives from the terminal abdominal ganglion of freely moving wood crickets (Nemobius sylvestris) in a semi-field situation. A cluster analysis of spike amplitudes revealed six clusters, or 'units', corresponding to six different subsets of cercal interneurons. No spontaneous activity was recorded for the units of larger amplitude, reinforcing the idea they correspond to the largest giant interneurons. Many of the cercal units are already activated by background noise, sometimes only weakly, and the approach of a predator is signaled by an increase in their activity, in particular for the larger-amplitude units. A scaling law predicts that the cumulative number of spikes is a function of the velocity of the flow perceived at the rear of the cricket, including a multiplicative factor that increases linearly with piston velocity. We discuss the implications of this finding in terms of how the cricket might infer the imminence and nature of a predatory attack.

Encoding of stimulus velocity by insect cercal system wind-sensitive interneurons: A comparative study.

Encoding of stimulus velocity by insect cercal system wind-sensitive interneurons: A comparative study.

Danger detection and escape behaviour in wood crickets

The wind-sensitive cercal system of Orthopteroid insects that mediates the detection of the approach of a predator is a very sensitive sensory system. It has been intensively analysed from a behavioural and neurobiological point of view, and constitutes a classical model system in neuroethology. The escape behaviour is triggered in orthopteroids by the detection of air-currents produced by approaching objects, allowing these insects to keep away from potential dangers. Nevertheless, escape behaviour has not been studied in terms of success. Moreover, an attacking predator is more than “air movement”, it is also a visible moving entity. The sensory basis of predator detection is thus probably more complex than the perception of air movement by the cerci. We have used a piston mimicking an attacking running predator for a quantitative evaluation of the escape behaviour of wood crickets Nemobius sylvestris. The movement of the piston not only generates air movement, but it can be seen by the insect and can touch it as a natural predator. This procedure allowed us to study the escape behaviour in terms of detection and also in terms of success. Our results showed that 5–52% of crickets that detected the piston thrust were indeed touched. Crickets escaped to stimulation from behind better than to a stimulation from the front, even though they detected the approaching object similarly in both cases. After cerci ablation, 48% crickets were still able to detect a piston approaching from behind (compared with 79% of detection in intact insects) and 24% crickets escaped successfully (compared with 62% in the case of intact insects). So, cerci play a major role in the detection of an approaching object but other mechanoreceptors or sensory modalities are implicated in this detection. It is not possible to assure that other sensory modalities participate (in the case of intact animals) in the behaviour; rather, than in the absence of cerci other sensory modalities can partially mediate the behaviour. Nevertheless, neither antennae nor eyes seem to be used for detecting approaching objects, as their inactivation did not reduce their detection and escape abilities in the presence of cerci.

The Cercal Organ May Provide Singing Tettigoniids a Backup Sensory System for the Detection of Eavesdropping Bats

Conspicuous signals, such as the calling songs of tettigoniids, are intended to attract mates but may also unintentionally attract predators. Among them bats that listen to prey-generated sounds constitute a predation pressure for many acoustically communicating insects as well as frogs. As an adaptation to protect against bat predation many insect species evolved auditory sensitivity to bat-emitted echolocation signals. Recently, the European mouse-eared bat species Myotis myotis and M. blythii oxygnathus were found to eavesdrop on calling songs of the tettigoniid Tettigonia cantans. These gleaning bats emit rather faint echolocation signals when approaching prey and singing insects may have difficulty detecting acoustic predator-related signals. The aim of this study was to determine (1) if loud self-generated sound produced by European tettigoniids impairs the detection of pulsed ultrasound and (2) if wind-sensors on the cercal organ function as a sensory backup system for bat detection in tettigoniids. We addressed these questions by combining a behavioral approach to study the response of two European tettigoniid species to pulsed ultrasound, together with an electrophysiological approach to record the activity of wind-sensitive interneurons during real attacks of the European mouse-eared bat species Myotis myotis. Results showed that singing T. cantans males did not respond to sequences of ultrasound pulses, whereas singing T. viridissima did respond with predominantly brief song pauses when ultrasound pulses fell into silent intervals or were coincident with the production of soft hemi-syllables. This result, however, strongly depended on ambient temperature with a lower probability for song interruption observable at 21°C compared to 28°C. Using extracellular recordings, dorsal giant interneurons of tettigoniids were shown to fire regular bursts in response to attacking bats. Between the first response of wind-sensitive interneurons and contact, a mean time lag of 860 ms was found. This time interval corresponds to a bat-to-prey distance of ca. 72 cm. This result demonstrates the efficiency of the cercal system of tettigoniids in detecting attacking bats and suggests this sensory system to be particularly valuable for singing insects that are targeted by eavesdropping bats.

Relative contributions of organ shape and receptor arrangement to the design of cricket’s cercal system

Understanding the relative contributions of the shape of a sensory organ and the arrangement of receptors to the overall performance of the organ has long been a challenge for sensory biologists. We tackled this issue using the wind-sensing system of crickets, the cerci, two conical abdominal appendages covered with arrays of filiform hairs. Scanning electron microscopy coupled with 3D reconstruction methods were used for mapping of all cercal filiform hairs. The hairs are arranged according to their diameter in a way that avoids collisions with neighbours during hair deflection: long hairs are regularly spaced, whereas short hairs are both randomly and densely distributed. Particle image velocimetry showed that the variation in diameter of the cercus along its length modifies the pattern of fluid velocities. Hairs are subject to higher air flow amplitudes at the base than at the apex of the cercus. The relative importance of interactions between receptors and the air flow around the organ may explain the performance of the cricket's cercal system: it is characterised by a high density of statistically non-interacting short hairs located at the base of the cercus where sensitivity to air currents is the highest.

NEUROSENSORY ECOLOGY IN THE JEB

It is often difficult to imagine how other people perceive the world, let alone other species, so when Ken Lukowiak was approached by The JEB's Editor in Chief, Hans Hoppeler, to draw together reviews discussing Neurosensory Ecology, he decided to steer clear of the usual sensory topics. Taking an

Free-flight encounters between praying mantids (Parasphendale agrionina) and bats (Eptesicus fuscus)

Through staged free-flight encounters between echolocating bats and praying mantids, we examined the effectiveness of two potential predator-evasion behaviors mediated by different sensory modalities: (1) power dive responses triggered by bat echolocation detected by the mantis ultrasound-sensitive auditory system, and (2) ;last-ditch' maneuvers triggered by bat-generated wind detected by the mantis cercal system. Hearing mantids escaped more often than deafened mantids (76% vs 34%, respectively; hearing conveyed 42% advantage). Hearing mantis escape rates decreased when bat attack sequences contained very rapid increases in pulse repetition rates (escape rates <40% for transition slopes >16 p.p.s. 10 ms(-1); escape rates >60% for transition slopes <16 p.p.s. 10 ms(-1)). This suggests that echolocation attack sequences containing very rapid transitions (>16 p.p.s. 10 ms(-1)) could circumvent mantis/insect auditory defenses. However, echolocation attack sequences containing such transitions occurred in only 15% of the trials. Since mantis ultrasound-mediated responses are not 100% effective, cercal-mediated evasive behaviors triggered by bat-generated wind could be beneficial as a backup/secondary system. Although deafened mantids with functioning cerci did not escape more often than deafened mantids with deactivated cerci (35% vs 32%, respectively), bats dropped mantids with functioning cerci twice as frequently as mantids with deactivated cerci. This latter result was not statistically reliable due to small sample sizes, since this study was not designed to fully evaluate this result. It is an interesting observation that warrants further investigation, however, especially since these dropped mantids always survived the encounter.

8.5. A neuroanatomical guide of the cercal scape system of the wood cricket

8.5. A neuroanatomical guide of the cercal scape system of the wood cricket

Construction of Point Process Adaptive Filter Algorithms for Neural Systems Using Sequential Monte Carlo Methods

The stochastic state point process filter (SSPPF) and steepest descent point process filter (SDPPF) are adaptive filter algorithms for state estimation from point process observations that have been used to track neural receptive field plasticity and to decode the representations of biological signals in ensemble neural spiking activity. The SSPPF and SDPPF are constructed using, respectively, Gaussian and steepest descent approximations to the standard Bayes and Chapman-Kolmogorov (BCK) system of filter equations. To extend these approaches for constructing point process adaptive filters, we develop sequential Monte Carlo (SMC) approximations to the BCK equations in which the SSPPF and SDPPF serve as the proposal densities. We term the two new SMC point process filters SMC-PPFs and SMC-PPFD, respectively. We illustrate the new filter algorithms by decoding the wind stimulus magnitude from simulated neural spiking activity in the cricket cercal system. The SMC-PPFs and SMC-PPFD provide more accurate state estimates at low number of particles than a conventional bootstrap SMC filter algorithm in which the state transition probability density is the proposal density. We also use the SMC-PPFs algorithm to track the temporal evolution of a spatial receptive field of a rat hippocampal neuron recorded while the animal foraged in an open environment. Our results suggest an approach for constructing point process adaptive filters using SMC methods.

Ontogeny of air-motion sensing in cricket

Juvenile crickets suffer high rates of mortality by natural predators that they can detect using extremely sensitive air-sensing filiform hairs located on their cerci. Although a huge amount of knowledge has accumulated on the physiology, the neurobiology and the biomechanics of this sensory system in adults, the morphological and functional aspects of air sensing have not been as well studied in earlier life history stages. Using scanning electronic microscopy, we performed a survey of all cercal filiform hairs in seven instars of the wood cricket (Nemobius sylvestris). Statistical analyses allowed us to quantify profound changes in the number, the length and the distribution of cercal hairs during development. Of particular importance, we found a fivefold increase in hair number and the development of a bimodal length-frequency distribution of cercal hairs from the second instar onwards. Based on theoretical estimations of filiform hair population coding, we found that the cercal system is functional for a wide range of frequencies of biologically relevant oscillatory flows, even from the first instar. As the cricket develops, the overall sensitivity of the cercal system increases as a result of the appearance of new hairs, but the value of the best tuned frequency remains fixed between 150 and 180 Hz after the second instar. These frequencies nicely match those emitted by natural flying predators, suggesting that the development of the cercal array of hairs may have evolved in response to such signals.

Co-factors and co-repressors of Engrailed: expression in the central nervous system and cerci of the cockroach, Periplaneta americana

In the larval cockroach (Periplaneta americana), knockout of Engrailed (En) in the medial sensory neurons of the cercal sensory system changes their axonal arborization and synaptic specificity. Immunocytochemistry has been used to investigate whether the co-repressor Groucho (Gro; vertebrate homolog: TLE) and the co-factor Extradenticle (Exd; vertebrate homolog: Pbx) are expressed in the cercal system. Gro/TLE is expressed ubiquitously in cell nuclei in the embryo, except for the distal pleuropodia. Gro is expressed in all nuclei of the thoracic and abdominal central nervous system (CNS) of first instar larva, although some neurons express less Gro than others. Cercal sensory neurons express Gro protein, which might therefore act as a co-repressor with En. Exd/Pbx is expressed in the proximal portion of all segmental appendages in the embryo, with the exception of the cerci. In the first instar CNS, Exd protein is expressed in subsets of neurons (including dorsal unpaired medial neurons) in the thoracic ganglia, in the first two abdominal ganglia, and in neuromeres A8-A11 of the terminal ganglion. Exd is absent from the cerci. Because Ultrabithorax/Abdominal-A (Ubx/Abd-A) can substitute for Exd as En co-factors in Drosophila, Ubx/Abd-A immunoreactivity has also been investigated. Ubx/Abd-A immunostaining is present in abdominal segments of the embryo and first instar CNS as far caudal as A7 and faintly in the T3 segment. However, Ubx/Abd-A is absent in the cerci and their neurons. Thus, in contrast to its role in Drosophila segmentation, En does not require the co-factors Exd or Ubx/Abd-A in order to control the synaptic specificity of cockroach sensory neurons.

Wind generated by an attacking bat: anemometric measurements and detection by the praying mantis cercal system

The wind-sensitive cercal system, well-known for mediating terrestrial escape responses, may also mediate insect aerial bat-avoidance responses triggered by wind generated by the approaching bat. One crucial question is whether enough time exists between detection and capture for the insect to perform a successful evasive maneuver. A previous study estimated this time to be 16 ms, based on cockroach behavioral latencies and a prediction for the detection time derived from a simulated predator moving toward a simulated prey. However, the detection time may be underestimated since both the simulated predator and prey lacked certain characteristics present in the natural situation. In the present study, actual detection times are measured by recording from wind-sensitive interneurons of a tethered praying mantis that serves as the target for a flying, attacking bat. Furthermore, using hot-wire anemometry, we describe and quantify the wind generated by an attacking bat. Anemometer measurements revealed that the velocity of the bat-generated wind consistently peaks early with a high acceleration component (an important parameter for triggering wind-mediated terrestrial responses). The physiological recordings determined that the mantis cercal system detected an approaching bat 74 ms before contact, which would provide the insect with 36 ms to perform a maneuver before capture. This should be sufficient time for the mantis to respond. Although it probably would not have time for a full response that completely evades the bat, even a partial response might alter the mantid's trajectory enough to cause the bat to mishandle the insect, allowing it to escape.

Dejittered spike-conditioned stimulus waveforms yield improved estimates of neuronal feature selectivity and spike-timing precision of sensory interneurons.

What is the meaning associated with a single action potential in a neural spike train? The answer depends on the way the question is formulated. One general approach toward formulating this question involves estimating the average stimulus waveform preceding spikes in a spike train. Many different algorithms have been used to obtain such estimates, ranging from spike-triggered averaging of stimuli to correlation-based extraction of "stimulus-reconstruction" kernels or spatiotemporal receptive fields. We demonstrate that all of these approaches miscalculate the stimulus feature selectivity of a neuron. Their errors arise from the manner in which the stimulus waveforms are aligned to one another during the calculations. Specifically, the waveform segments are locked to the precise time of spike occurrence, ignoring the intrinsic "jitter" in the stimulus-to-spike latency. We present an algorithm that takes this jitter into account. "Dejittered" estimates of the feature selectivity of a neuron are more accurate (i.e., provide a better estimate of the mean waveform eliciting a spike) and more precise (i.e., have smaller variance around that waveform) than estimates obtained using standard techniques. Moreover, this approach yields an explicit measure of spike-timing precision. We applied this technique to study feature selectivity and spike-timing precision in two types of sensory interneurons in the cricket cercal system. The dejittered estimates of the mean stimulus waveforms preceding spikes were up to three times larger than estimates based on the standard techniques used in previous studies and had power that extended into higher-frequency ranges. Spike timing precision was approximately 5 ms.