The effects of transplantation and regeneration of sensory neurons on a somatotopic map in the cricket central nervous system

The restoration of the cercal afferent projection of crickets was examined after severing the cercal nerve or amputating the cercus and reimplanting it. After either maneuver the sensory neurons regenerated arborizations in the central nervous system (CNS) within about 1 month. In order to assess the role of the pathway taken to the CNS in controlling the growth of the terminal arborization, we transplantated left cerci to the right side of the host. The operation mismatched the mediolateral axes of host and graft tissues. In one-third of the neurons examined, the axon trajectories of the regenerated neurons were altered. The terminal arborizations in these cases were unusual; for example, one neuron arborized in an abnormal area as well as in its normal area. In rare instances this neuron arborized only in incorrect areas of the CNS. Thus, it appears that axon pathway can have an effect on the central structure of sensory neurons. However, in most cases after the surgery, the neurons were able to reach their proper target areas even by circuitous routes. The proximodistal coordinate of the map is isomorphic with sensory neuron age, because the most distal receptors are produced early in postembryonic development and new ones are added proximally at each molt. We tested the possibility that the order of differentiation was critical for generating the afferent projection with two experiments. First, the distal cercus including the distal members of the clavate array was amputated. The specimen regenerated an entire distal cercus including distal clavate receptors. When newly generated, distal neurons were stained, the terminal arbors were identical to the amputated neurons they replaced. In this case, both age and order of arrival were reversed from normal yet the topographic projection pattern was not altered. Second, we transplanted young cerci onto older specimens and then examined the regenerated arbors of the transplanted sensory neuron. The immature neuron arborized in the adult nervous system exactly as the mature homolog. Thus the age of a sensory neuron did not appear to be a controlling variable in the elaboration of a terminal arborization. The significance of these results is discussed in the context of two models for development of orderly neuronal connections.     

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.     

Tracheation of abdominal ganglia and cerci in the house cricket Acheta domesticus (Orthoptera,Gryllidae)

Patterns of tracheation in the abdominal central nervous system and the cerci of Acheta domesticus are described from whole mounts, and light and electron microscopy. The tracheal supply of the ganglia is derived from ventral longitudinal tracheal trunks which have segmental connections to the spiracels. Each abdominal ganglion is served by a single pair of tracheal trunks, except the terminal ganglion, which has two pairs. Within the ganglia, tracheoles occur principally in association with glia-rich areas of the neuropile. We suggest that the respiratory exchange may be concentrated in the cell bodies of neurons and glia. Each cercus has a tracheal supply in paralle with a large air sac which, it is suggested, serves to lighten the cercus, functions as a resonator for sound reception, or facilitates tidal flow of hemolymph and postecdysial expansion of the cercus. No tracheae run continuously between ganglia or between the terminal ganglion and the cerci, and they do not appear to have a potential role as a contact guidance pathway for cercal nerve growth.     

Effect of body orientation in the gravitational field on directional sensitivity of the cercal system of neurons in crickets

Changes in the spatial orientation of three-dimensional directional sensitivity diagrams of neurons of the terminal abdominal ganglion of the cricket during body tilting were studied. Spike responses were recorded from neurons of the ganglion to acoustic stimuli in different directions, with the cricket's body tilted at different angles to the horizontal plane. During tilting of the cricket's body the orientation of the directional sensitivity diagrams was found to change parallel with the orientation of the body. Neurons of the abdominal ganglion are excited by cercal sensillae, among which there are receptors which respond to changes in the position of the cricket's body in the gravitational field (gravity receptors). The results suggest that cercal gravity receptors have no specific influence on the directional sensitivity of neurons of the first central division of the cercal system.Institute for Problems in Information Transmission, Academy of Sciences of the USSR, Moscow. Translated from Neirofiziologiya, Vol. 12, No. 6, pp. 604–611, November–December, 1980.     

Constant representation of signal spatial location in the cricket cercal system]

Directional sensitivity of the abdominal giant fibers of the cricket to auditory stimulation has been investigated with special interest to the effect of the cerci position on the directional sensitivity of the giant neurons of the terminal abdominal ganglion. It has been shown that at least for some giant neurons the preferred directions of stimulation are practically independent of cerci position. (Fig. 2, 3). By other words these neurons have constant preferred directions in relation to the insect body, but not to the cerci. This property of giant neurons can be accounted for their changeable connections with many receptors having different directional selectivity. If innervation of the giant neuron is artificially restricted to one group of receptors with identical directional selectivity the described constancy of preferred directions in relation to the body disappears (Fig. 4).     

Synapse formation by sensory neurons after cross-species transplantation in crickets: the role of positional information

The role of positional information in synapse formation was studied in the cricket cercal sensory system by transplanting epidermis from one species of cricket to another. Strips of cercal epidermis containing identified sensory neurons were transplanted from a black donor species to a tan host species; the color difference was used to distinguish between donor and host tissue in adults. Transplanted sensory neurons regenerated axons into the host terminal abdominal ganglion where they formed functional chimeric synapses. These methods were used to test the role of positional information in central synapse formation. Newly generated sensory neurons, formed by the donor tissue at the border between graft and host, were examined to test the idea that their position would determine their structure, function, and projection pattern. These "intercalated" sensory neurons support the positional information hypothesis. First, they had directional sensitivities which were appropriate to their location on the cercus; receptors of this directionality would never be made by the donor tissue if left in its original position. Second, these sensory neurons projected to regions of the CNS known to be appropriate for their directionality. Finally, simultaneous recordings from these ectopic sensory neurons and host interneurons demonstrated the expected synaptic connection, based on the overlap of pre- and postsynaptic cells. Thus three aspects of receptor function, directionality, afferent projection, and choice of synaptic partners, appeared to be controlled by positional information.     

Innervation of cockroach cercal receptors as revealed by horseradish peroxidase (Insecta,Blattodea)

Summary The innervation of cerci of a desert burrowing cockroach, Arenivaga sp., was determined by horseradish peroxidase backfilling of the cercal nerve and histochemistry. The procedure yielded a high percentage of successful fills and in many cases every neuron filled completely, including dendrites and axons of less than one m. The innervation of the cerci was found to be highly ordered. Upon entering the cercus, the cercal nerve splits into bilateral branches, one on each side of the midline. The nerves branch again at each segment to form fascicles of sensory neurons which innervate the trichobothria, sensilla chaetica and tricholiths, each with a single bipolar neuron. While the cell bodies of neurons are of similar dimensions, the dendrites to the tricholiths are much longer and terminate on the midline side of the sensilla socket where the tricholith shaft attaches.     

Cercal sensory system and giant interneurons in Gryllodes sigillatus

The external morphologies of two cricket species, Gryllodes sigillatus and Gryllus bimaculatus, were investigated. Despite its small body length, G. sigillatus possessed longer cerci and longer cercal filiform hairs than G. bimaculatus. The estimated number of filiform hairs on a cercus was also larger in G. sigillatus than in G. bimaculatus. Wind-sensitive interneurons receiving sensory inputs from cercal filiform hairs and running in the ventral nerve cord (VNC) were investigated in G. sigillatus both morphologically and physiologically. By intracellular staining, these interneurons were proved to be morphologically homologous with previously identified giant interneurons (GIs 8-1, 9-1, 9-2, 9-3, 10-2, and 10-3) in G. bimaculatus and Acheta domesticus. In G. sigillatus, the intensity-response relationship (I-R curve) for each GI was investigated using a unidirectional air current stimulus. The stimulus was applied from 12 different directions, and an I-R curve was obtained for each stimulus direction. Each GI showed a characteristic I-R curve depending on stimulus direction. The directionality curve expressed in terms of threshold velocity showed that each GI had a distinctive directional characteristic. The functional properties of GIs in G. sigillatus, such as I-R curve, threshold velocity, and directional characteristics, were compared with those of homologous GIs in G. bimaculatus in Discussion.     

Evolution of the cercal sensory system in a tropical cricket clade (Orthoptera: Grylloidea: Eneopterinae): a phylogenetic approach

The diversity of sensory systems in animals has poorly been explored on a phylogenetic basis at the species level. We addressed this issue using cricket cerci, comprising abdominal appendages covered with touch‐ and air‐sensitive hairs. Scanning electron microscopy measurements and spatial analyses of hair positioning were used to quantify the structural diversity of cercal structures. Eighteen Eneopterinae and two Gryllidae (outgroups) were studied from a phylogenetic perspective. Cerci were revealed to be complex, diverse, and variable between cricket species. Based on maximum likelihood estimations, the ancestral Eneopterinae cercus had a small size, and its hair equipment allowed the use of both air and touch mechanoreception. The evolution of Eneopterinae cerci was mainly unconstrained by the phylogeny; it was rather a punctuated process, involving apical transformations, and was mostly unrelated to environmental patterns. All studied species have enhanced their overall perceptive capacities compared to the ancestor. Most have longer cerci with more and/or longer hairs. Sensory abilities have improved either in the direction of touch or air movement detection, or both, without discarding the potential for any sensory capacity that was already present ancestrally. This pattern is consistent with the hypothesis of an evolutionary trade‐off for sensory performances. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 99 , 614–631.     

Direction sensitivity of the filiform hair population of the cricket cereal system

Each cercus on the adult cricket Acheta domesticus bears 1000–2000 filiform hair mechanoreceptors. In order to determine the extent of identifiability of individual hair receptors, the structural characteristics of ten putative identified hairs were measured in 21–25 different animals. For these ten hairs, the sets of preferred directions and circumferential locations had standard deviations of 6.8° and 5.9°, respectively. There was no significant inter-animal covariance between a hair's preferred direction and its circumferential location. The preferred directions of 246 different identified hairs were then measured from 16 animals in order to characterize the distribution of preferred directions of hairs on a single typical cercus. These data were transformed from the frame of reference of the cercus into that of the cricket, generating an estimate of the representation of air-current stimulus direction provided by the entire ensemble of filiform hairs on both cerci. The distribution of hair directional sensitivities was continuous but extremely non-uniform, and more complex than previous studies had suggested.Abbreviations MT medial-transverse - LT lateral-transverse - AL anterior-longitudinal - PL posterior-longitudinal - MAO medial-anterior-oblique - MPO medial-posterior-oblique - LAO lateral-anterior-oblique - LPO lateral-posterior-oblique - T transverse - L longitudinal - O oblique - CNS central nervous system     

The cercal receptor system of the praying mantid,Archimantis brunneriana Sauss.

Summary The cerci of the praying mantid, Archimantis brunneriana Sauss., are paired segmented sensory organs located at the tip of the abdomen. Basally the cercal segments are slightly flattened dorso-ventrally and are fused to such a degree that it is difficult to distinguish them. Distally the segments become progressively more flattened laterally and their boundaries become more obvious.Two types of sensilla are present on the cerci, trichoid sensilla and filiform sensilla. Trichoid hairs are longest on the medial side of the cerci and toward the cercal base. On the proximal cercal segments they are grouped toward the middle of each segment while they are more uniformly distributed on the distal segments. Filiform sensilla are found at the distal end of each segment except the last and are most abundant on the middle segments of the cercus. Both the number of cercal segments and the number of sensilla are variable. Trichoid hairs are highly variable in appearance from short and stout to long and thin. They arise from a raised base, have a fluted shaft, and some have a pore at the tip. They are innervated by from one to five dendrites, one of which is always considerably larger than the others. Some of the dendrites continue out into the shaft of the hair.Filiform hairs have fluted shafts and are mounted in a flexible membrane within a cuticular ring in a depression. They are innervated by a single large sensory neuron, the dendrite of which passes across a flattened area on the inner wall of the lumen of the hair. The dendritic sheath forms the lining of the ecdysial canal and is therefore firmly attached to the hair. The dendrite is attached to the sheath by desmosomes distally and is penetrated by projections of the sheath more proximally. A fibrous cap surrounds the dendrite and may hold it in place relative to the hair.The cercal receptor system of Archimantis is compared to those of cockroaches and crickets.     

Regeneration of cercal filiform hair sensory neurons in the first-instar cockroach restores escape behavior

Neural regeneration in the escape circuit of the first-instar cockroach is described using behavioral analysis, electrophysiology, intracellular staining, and electron microscopy. Each of the two filiform hairs on each of the animal's cerci is innervated by a single sensory neuron, which specifically synapses with a set of giant interneurons (GIs) in the terminal ganglion. These trigger a directed escape run. Severing the sensory axons causes them to degenerate and perturbs escape behavior, which is restored to near normal after 4–6 days. Within this time, afferents regenerate and reestablish arborizations in the terminal ganglion. In most cases, regenerating afferents enter the cercal glomerulus and re-form most of the specific monosynaptic connections they acquired during embryogenesis, although their morphology deviates markedly from normal; these animals reestablish near normal escape behavior. In a few cases, regenerating afferents remain within the cercus or bypass the cercal glomerulus, and thereby fail to re-form synapses with GIs; these animals continue to exhibit perturbed escape behavior. We conclude that in most cases, specific synapses are reestablished and appropriate escape behavior is restored. This regeneration system therefore provides a tractable model for the establishment of synaptic specificity in a simple neuronal circuit. © 1997 John Wiley & Sons, Inc. J Neurobiol 33: 439–458, 1997     

Visualization of ensemble activity patterns of mechanosensory afferents in the cricket cercal sensory system with calcium imaging

The cercal sensory system of the cricket mediates the detection and analysis of low velocity air currents in the animal's immediate environment, and is implemented around an internal representation of air current direction that demonstrates the essential features of a continuous neural map. Previous neurophysiological and anatomical studies have yielded predictions of the global spatio-temporal patterns of activity that should be evoked in the sensory afferent map by air current stimuli of different directions. We tested those predictions by direct visualization of ensemble afferent activity patterns using Ca2+ -sensitive indicators. The AM ester of the fluorescent Ca2+ indicator (Oregon Green 488 BAPTA-1 AM) was injected under the sheath of a cercal sensory nerve containing all of the mechanosensory afferent axons from one cercus. Optical signals were recorded with a digital intensified CCD camera. Control experiments using direct electrical stimulation of stained and unstained nerves demonstrated that the observed Ca2+ signals within the terminal abdominal ganglion (TAG) were due to activation of the dye-loaded sensory afferent neurons. To visualize the spatial patterns of air-current-evoked ensemble activity, unidirectional air currents were applied repeatedly from eight different directions, and the optically recorded responses from each direction were averaged. The dispersion of the optical signals by the ganglion limited the spatial resolution with which these ensemble afferent activity patterns could be observed. However, resolution was adequate to demonstrate that different directional stimuli induced different spatial patterns of Ca2+ elevation in the terminal arbors of afferents within the TAG. These coarsely- resolved, optically-recorded patterns were consistent with the anatomy-based predictions.     

Interaction between arthropod filiform hairs in a fluid environment

Many arthropods use filiform hairs as mechanoreceptors to detect air motion. In common house crickets (Acheta domestica) the hairs cover two antenna-like appendages called cerci at the rear of the abdomen. The biomechanical stimulus-response properties of individual filiform hairs have been investigated and modeled extensively in several earlier studies. However, only a few previous studies have considered viscosity-mediated coupling between pairs of hairs, and only in particular configurations. Here, we present a model capable of calculating hair-to-hair coupling in arbitrary configurations. We simulate the coupled motion of a small group of mechanosensory hairs on a cylindrical section of cercus. We have found that the coupling effects are non-negligible, and likely constrain the operational characteristics of the cercal sensory array.     

A gradient of synaptic efficacy and its presynaptic basis in the cercal system of the cockroach

1. The cerci of the cockroach Periplaneta americana bear longitudinal columns of wind-sensitive receptors which provide excitatory inputs to the giant interneurons (GIs) of the abdominal nerve cord. By using sound stimuli, we showed that spikes were more easily induced in the GIs from the most proximal than from the most distal receptors of the same column. 2. This was not due to a greater responsiveness of proximal sensilla to tones but to stronger synaptic connections; for the 3 largest GIs, the amplitude of the monosynaptic unitary EPSP tended to be all the higher as the stimulated sensillum was more proximal in each column. 3. The differences in EPSP size were due, at least partly, to presynaptic factors: a statistical analysis of the amplitude fluctuations of single-fibre EPSPs, showed that the amount of transmitter released per presynaptic impulse was larger for proximal than for distal sensory neurons in each column. 4. These differences in synaptic strength were correlated with differences in the structure of the afferent terminals. The location, the size and the shape of the axonal arbors are nearly the same for all sensory neurons of the same column, but proximal neurons arborize more profusely, and the terminal arbor of distal neurons is generally characterized by dorsal clusters of varicosities. 5. During postembryonic development, a decrease in the connection strength of 2 identified cercal neurons was accompanied by a retraction of ramifications on the medial side of their axonal arbor. 6. Possible mechanisms involved in the genesis and the remodelling of the gradient of synaptic strength are discussed in the light of available data and hypotheses relative to the development of ordered afferent connections.     

Synchronous firing by specific pairs of cercal giant interneurons in crickets encodes wind direction

One prominent stimulus to evoke an escape response in crickets is the detection of air movement, such as would result from an attacking predator. Wind is detected by the cercal sensory system that consists of hundreds of sensory cells at the base of filiform hairs. These sensory cells relay information to about a dozen cercal giant and non-giant interneurons. The response of cercal sensory cells depends both, on the intensity and the direction of the wind. Spike trains of cercal giant interneurons then convey the information about wind direction and intensity to the central nervous system. Extracellular recording of multiple cercal giant interneurons shows that certain interneuron pairs fire synchronously if a wind comes from a particular direction. We demonstrate here that directional tuning curves of synchronously firing pairs of interneurons are sharper than those of single interneurons. Moreover, the sum total of all synchronously firing pairs eventually covers all wind directions. The sharpness of the tuning curves in synchronously firing pairs results from excitatory and inhibitory input from the cercal sensory neurons. Our results suggest, that synchronous firing of specific pairs of cercal giant interneurons encodes the wind direction. This was further supported by behavioral analyses.     

Two types of information processing in cercal systems of insects: directional sensitivity of giant interneurons

Summary Cercal systems of seven insect species (cricketMelanogryllus desertus, mole cricketGryllotalpa gryllotalpa, katydidsPholidoptera pustulipes andTettigonia viridissima, cockroachesPeriplaneta americana andBlatta orientalis, and locustLocusta migratoria) were examined for direction-sensitive giant interneurons (GIs) that are excited by cercal receptors but have directional preferences independent of cercus position. Such GIs are known for the cricketsAcheta domesticus andGryllus bimaculatus. Directional sensitivity diagrams (DSDs) of GIs were obtained by recording and analysing the electrical responses of abdominal connectives to sound stimuli from various directions. For each animal DSDs were plotted in the form of polar graphs for two or three positions of the stimulated cercus so that the effect of cercus position on the orientation of the DSD could be evaluated.All insects studied had GIs whose DSDs for fixed cercus positions were similar in appearance to the DSDs described for GIs of the cricketsAcheta domesticus andGryllus bimaculatus. Most of these DSDs were shaped like a figure 8 (when airflow is used as the stimulus instead of sound, each DSD has only one lobe). However, not all GIs demonstrated a constant directional preference. GIs with constant directionality were found only inMelanogryllus desertus, Pholidoptera pustulipes, Tettigonia viridissima andLocusta migratoria. In these insects DSDs from one GI plotted for different cercus positions had approximately the same orientation (Figs. 4–7). In contrast, GIs inGryllotalpa gryllotalpa, Periplaneta americana andBlatta orientalis had DSDs whose orientation changed in accordance with a change in position of the stimulated cercus (Figs. 8–10).Thus, direction-sensitive GIs investigated here can be divided into two types: (1) GIs with constant directionality (whose DSDs are fixed to the body, and (2) GIs with variable directionality (whose DSDs are fixed to the cerci). To date, in each species only GIs of the same type have been encountered. This may be an indication that cercal systems can be divided into two categories according to how they process information. However, since we have not tested all GIs in each species, we cannot rule out the possibility that a species might have both types of GIs.Abbreviations DSD directional sensitivity diagram - GI giant interneuron - TAG terminal abdominal ganglion     

Ectopic neurons and the organization of insect sensory systems

Summary The transplantation of appendages from one place to another on the body of crickets (Acheta domesticus) has been used to study the similarities and differences between the sensory systems of various ganglia. Mesothoracic legs have been transplanted to the abdomen in place of a cercus and cerci have been transplanted to thoracic leg stumps. After the ectopic sensory neurons had time to regenerate into the CNS, they were stained and their axonal arborizations examined. The results, which were concerned primarily with bristle receptors, revealed that bristle afferents on ectopic cerci arborized in ventral neuropil (the ventralmost association center) and leg afferents arborized in a ventral anterior region of the terminal abdominal ganglion. The results support the idea that each ganglion contains only a few distinct regions of neuropil (probably three), each receiving separate subsets of the afferent projection.The ectopic cerci were also shown to excite interneurons in the thoracic ganglia whose dendrites were located in the most ventral neuropil. These neurons normally respond to thoracic bristle afferents. Thus, the segregation of afferent axons has a correlate in the interneurons they excite.     

Quantitative characterization of the filiform mechanosensory hair array on the cricket cercus

Background

Crickets and other orthopteran insects sense air currents with a pair of abdominal appendages resembling antennae, called cerci. Each cercus in the common house cricket Acheta domesticus is approximately 1 cm long, and is covered with 500 to 750 filiform mechanosensory hairs. The distribution of the hairs on the cerci, as well as the global patterns of their movement vectors, have been characterized semi-quantitatively in studies over the last 40 years, and have been shown to be very stereotypical across different animals in this species. Although the cercal sensory system has been the focus of many studies in the areas of neuroethology, development, biomechanics, sensory function and neural coding, there has not yet been a quantitative study of the functional morphology of the receptor array of this important model system.

Methodology/Principal Findings

We present a quantitative characterization of the structural characteristics and functional morphology of the cercal filiform hair array. We demonstrate that the excitatory direction along each hair''s movement plane can be identified by features of its socket that are visible at the light-microscopic level, and that the length of the hair associated with each socket can also be estimated accurately from a structural parameter of the socket. We characterize the length and directionality of all hairs on the basal half of a sample of three cerci, and present statistical analyses of the distributions.

Conclusions/Significance

The inter-animal variation of several global organizational features is low, consistent with constraints imposed by functional effectiveness and/or developmental processes. Contrary to previous reports, however, we show that the filiform hairs are not re-identifiable in the strict sense.     

Hair canopy of cricket sensory system tuned to predator signals

Filiform hairs located on the cerci of crickets are among the most sensitive sensors in the animal world and enable crickets to sense the faintest air movements generated by approaching predators. While the neurophysiological and biomechanical aspects of this sensory system have been studied independently for several decades, their integration into a coherent framework was wanting. In order to evaluate the hair canopy tuning to predator signals, we built a model of cercal population coding of oscillating air flows by the hundreds of hairs on the cerci of the sand cricket Gryllus bimaculatus (Insecta: Orthoptera). A complete survey of all hairs covering the cerci was done on intact cerci using scanning electronic microscopy. An additive population coding of sinusoid signals of varying frequencies and velocities taking into account hair directionality delivered the cercal canopy tuning curve. We show that the range of frequencies and velocities at which the cricket sensory system is best tuned corresponds to the values of signals produced by approaching predators. The relative frequencies of short (< 0.5 x 10(-3) m) and long hairs and their differing responses to oscillating air flows therefore enable crickets to detect predators in a time-frequency-intensity space both as far as possible and at close range.