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.     

Ectopic sensory neurons in mutant cockroaches compete with normal cells for central targets.

The cercus of the first instar cockroach, Periplaneta americana, bears two filiform hairs, lateral (L) and medial (M), each of which is innervated by a single sensory neuron. These project into the terminal ganglion of the CNS where they make synaptic connections with a number of ascending interneurons. We have discovered mutant animals that have more hairs on the cercus; the most typical phenotype, called "Space Invader" (SI), has an extra filiform hair in a proximo-lateral position on one of the cerci. The afferent neuron of this supernumerary hair (SIN) "invades the space" occupied by L in the CNS and makes similar synaptic connections to giant interneurons (GIs). SIN and L compete for these synaptic targets: the size of the L EPSP in a target interneuron GI3 is significantly reduced in the presence of SIN. Morphometric analysis of the L afferent in the presence or absence of SIN shows no anatomical concomitant of competition. Ablation of L afferent allows SIN to increase the size of its synaptic input to GI3. Less frequently in the mutant population, we find animals with a supernumerary medical (SuM) sensillum. Its afferent projects to the same neuropilar region as the M afferent, makes the same set of synaptic connections to GIs, and competes with M for these synaptic targets. The study of these competitive interactions between identified afferents and identified target interneurons reveals some of the dynamic processes that go on in normal development to shape the nervous system.     

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     

The terminal abdominal ganglion of the wood cricket Nemobius sylvestris

The abdominal cerci of the wood cricket, Nemobius sylvestris, are covered by a variety of hair‐like sensilla that differ in length, thickness, and articulation. Fillings from the cercal nerves with cobalt chloride and fluorescent dyes revealed the projection of sensory axons into the terminal abdominal ganglion of the ventral nerve chain. Two projection areas on each side of the terminal abdominal ganglion midline could be identified: a posterior cercal glomerulus and an anterior bristle neuropil. Axons from some cercal sensilla ascend through the connectives to reach the metathoracic ganglionic mass. As their axons pass through each segmental abdominal ganglion, they project medial arborization. Cross‐sections of the terminal abdominal ganglion and retrograde fills with cobalt chloride and fluorescent dyes from connectives revealed several small cells and seven pairs of giant ascending interneurons organized symmetrically. Giant somata are located contralateral to their axons (diameters between 20 and 45 μm). The cercal projections overlap extensively with the dendritic fields of the giant interneurons. In the terminal abdominal ganglion, we identified nine longitudinal tracts, two major tracts, and seven smaller ones. The functional implications of the neuranatomical organization of the system are discussed on a comparative basis. J. Morphol., 2008. © 2008 Wiley‐Liss, Inc.     

Positional information determines the anatomy and synaptic specificity of cockroach filiform hair afferents using independent mechanisms

Summary Mutant first instar cockroaches (Periplaneta americana) with supernumerary filiform hair sensilla on their cerci were used to study the effects of cell body position on axonal morphology and synaptic connections. The wild-type cercus has two hairs, one lateral (L) and the other medial (M), each with an underlying sensory neuron. Silver-intensified cobalt fills show that the supernumerary lateral neuron (SIN) in the mutant has the same shape of arborization as L, and electrophysiological recording shows that it forms synaptic connections with the same subset of giant interneurons (GIs) as L in the terminal ganglion: GI3 and GI6. The supernumerary medial neuron (SuM) has the same axonal morphology as M and synapses with the same GIs as does M: ipsilateral GIs 1 and 2 and contralateral GIs 1, 2, 3, 5 and 6. In 0.1% of approximately 8000 animals screened, a supernumerary hair arose on the cereal midline (C hair). The C neuron sends its axon to the CNS in the same branch of the cereal nerve as the L and SIN, and has a similar arborization. However, the C neuron forms synapses with the same GIs as do M and SuM. Electron microscopy of horseradish peroxidase-injected neurons was used to confirm that the C afferent forms a monosynaptic connection to GI2. It was concluded that the position of the sensory neuron cell body does control its axonal morphology and synaptic connectivity, but that these characteristics are produced by independent mechanisms.Abbreviations GI giant interneuron - L lateral - M medial - SI Space Invader - SuM supernumerary medial - C cereal midline     

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.     

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.     

Anatomy of equilibrium receptors and cerci of the burrowing desert cockroach Arenivaga (Insecta,Blattodea)

Summary The anatomy of the cerci of a burrowing desert cockroach, Arenivaga sp., with particular emphasis on equilibrium receptors was studied by light and scanning electron microscopy. Each cercus has two parallel rows of pendulous equilibrium receptors called tricholiths; the rows are parallel to the long axis of the cercus. The cerci and rows of tricholiths are orthogonally positioned with respect to the long axis of the insect at the posterior end of the abdomen and rotated laterally by about 25° from the horizontal plane. Tricholiths of each row are inserted in their sockets toward the midline of the cercus and their elliptical attachment to the gasket is parallel to the long axis of the cercus. The combination of these anatomical features constrains tricholith movements to 90° from the long axis of each cercus and accounts for the physiological findings that (1) interneurons receiving afference from tricholiths are maximally sensitive to displacements of the insect at 45°, 135°, 225°, and 315°, and (2) interneurons receiving input from the lateral rows respond to smaller displacements than those driven by medial rows (Walthall and Hartman 1981). First instar cockroaches have only one tricholith on each cercus. Subsequent molts result in the addition of tricholiths, usually paired. After six molts, the adult female bears six pairs; the adult male seven pairs of tricholiths. The additional afference is presumably required to drive the increasingly larger interneurons.     

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.     

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.     

Structure of the afferent terminals in terminal ganglion of a cricket and persistent homology

We use topological data analysis to investigate the three dimensional spatial structure of the locus of afferent neuron terminals in crickets Acheta domesticus. Each afferent neuron innervates a filiform hair positioned on a cercus: a protruding appendage at the rear of the animal. The hairs transduce air motion to the neuron signal that is used by a cricket to respond to the environment. We stratify the hairs (and the corresponding afferent terminals) into classes depending on hair length, along with position. Our analysis uncovers significant structure in the relative position of these terminal classes and suggests the functional relevance of this structure. Our method is very robust to the presence of significant experimental and developmental noise. It can be used to analyze a wide range of other point cloud data sets.     

Central projections of campaniform sensilla on the cerci of crickets and cockroaches

Summary Campaniform sensilla associated with filiform hairs comprise an important receptor type of the multimodal sensory system of the cerci of crickets and cockroaches. Their axon projections were investigated using iontophoretic cobalt injection into single sensilla.In crickets (Gryllus bimaculatus, Acheta domestica), six different types of cereal campaniform sensilla projections can be distinguished on the basis of their axonal arborizations and terminations. Typically, a proportion of cereal campaniform sensilla, associated with long filiform hairs, give rise to axons that ascend as through fibres from the terminal ganglion to reach the sixth abdominal ganglion. Cereal campaniform sensilla associated with clavate hairs have projections restricted to the terminal ganglion alone.Whereas in crickets axons of cercal campaniform sensilla invade only certain segmental neuropils in the terminal ganglion, in cockroaches (Periplaneta americana) axons from cercal campaniform sensilla branch in every segmental neuropil. A proportion of cereal campaniform sensilla in this species also gives rise to through fibres to the fifth abdominal ganglion.We discuss morphological and functional interpretations of differences between crickets and cockroaches and consider the significance of this type of receptor in the context of previous studies of the cercal system.     

Localization of the enhanced input to cockroach giant interneurons after partial deafferentation

The ventral giant interneurons (GIs) in the cockroach have two distinct dendritic fields: a small one ipsilateral to the soma, and a larger, contralateral field from which the axon arises. The major input to these GIs is from the cercus on the axon side; when this cercus is ablated in the last instar before the adult stage, input from the other cercus becomes more effective within 30 days (Vardi and Camhi, 1982b). I wished to determine if the input from the intact, soma-ipsilateral cercus contacted the GIs purely ipsilaterally and if EPSPs at this site were larger in deafferented animals. Consistent with earlier anatomical findings, intracellular recordings from the GI somata showed that the majority of cercal inputs synapse on their own side of the ganglion in normal animals. This was evidenced by differences in the size and shape of the synaptic potentials evoked from the two cerci and by the presence of large EPSPs after a ganglion had been split along the midline. Unitary EPSPs produced by stimulation of single, identified cercal afferents, ipsilateral to the soma, were compared between normal and deafferented animals. Column "h" afferents were chosen because they make a large contribution to the receptive fields of GIs 1 and 2 after ablation of the contralateral cercus. In addition, the arbors of these afferents, when stained with cobalt, did not cross the ganglionic midline in normal animals. Unitary EPSPs recorded in GI 2 were significantly larger in the deafferented animals. There was, however, no significant change in the size of EPSPs in GI 1. Nevertheless, the results from GI 2 suggest that partial deafferentation in the central nervous system can increase the efficacy of synapses distant from the locus of denervation.     

A second cricket cereal sensory system: bristle hairs and the interneurons they activate

Summary A new sensory system, in the abdomen of the cricketAcheta domesticus, is described. It consists of hair-like receptors, which we have called bristles, distributed on the cercus and abdomen. The sensory neurons, innervating bristles of a wide variety of shapes and sizes, project to a common area of the terminal abdominal ganglion. This region is distinct from the area called the cereal glomerulus which receives input from other receptor types.Three interneurons, whose dendrites are located exclusively in the projection area of bristle sensory neurons, are then described. These interneurons respond to tactile stimuli of the cercus and abdomen, but not to infrasound or to body orientation as do previously described interneurons. Based on the anatomical segregation of the afferents and interneurons, as well as the functional distinction from previously described cereal sensory systems, it is becoming clear that the cereal system is a multimodal sensory system.     

The morphology and fine structure of the giant interneurons of the wood cricket Nemobius sylvestris

The structural and ultrastructural characteristics of giant interneurons in the terminal abdominal ganglion of the cricket Nemobius sylvestris were investigated by means of cobalt and fluorescent dye backfilling and transmission electron microscopy.The projections of the 8 eight pairs of the biggest ascending interneurons (giant interneurons) are described in detail. The somata of all interneurons analyzed are located contralateral to their axons, which project to the posterior region of the terminal ganglion and arborise in the cercal glomerulus. Neuron 7-1a is an exception, because its arborisation is restricted to the anterior region of the ganglion. The fine structure of giant interneurons shows typical features of highly active cells. We observed striking indentations in the perineural layer, enabling the somata of the giant interneurons to be very close to the haemolymph. The cercal glomerulus exhibits a high diversity of synaptic contacts (i.e. axo-dendritic, axo-axonic, dendro-axonic, and dendro-dendritic), as well as areas of tight junctions. Electrical synapses seem to be present, as well as mixed synapses. The anatomical organization of the giant interneurons is finally discussed in terms of functional implications and on a comparative basis.     

Patterns of monosynaptic input to the giant interneurons 1–3 in the cereal system of the adult cockroach

1. The cerci of the cockroach Periplaneta americana bear filiform hair mechanoreceptors that are arranged in segmentally repeated rows and longitudinal columns. The monosynaptic connections between receptors of the same column or row and the 3 largest giant interneurons (GIs) were compared using the oil-gap single fibre technique.
2. For many columns, the synaptic efficacy of the afferents decreased gradually from the base to the tip of the cercus, but columns with an inverted gradient or without any gradient were also observed. On the ipsilateral side (relative to the GI axon), the inverted gradients were exclusively found for columns with short proximal hairs. For one column (d) and GI3, the ipsilateral and contralateral gradients were opposite.
3. Monosynaptic EPSPs evoked by stimulating different receptors of the same segment (segment 3) were of very different amplitudes, which partially account for the directional sensitivity of the GIs. Differences in the location, shape and size of the afferent terminals were not sufficient to explain these differences in connection strength.
4. No correlation was found between the size of the EPSPs produced by a sensory neuron and the length of its associated hair.

     

Long,smooth hair sensilla on the spider leg coxa: Sensory physiology,central projection pattern,and proprioceptive function (Arachnida,Araneida)

Summary Rows of long, smooth hair sensilla situated on both sides of the leg coxae were examined in the spider Cupiennius salei (Ctenidae). The hair shafts point into the space between adjacent legs and are deflected when the hairs of one coxa touch the cuticle of the neighboring coxa. 1. Unlike the serrated hair shafts of the ubiquitous tactile and chemosensitive setae of spiders, these hairs are entirely smooth. At their base they are articulated in a socket with an asymmetrical groove that determines the direction of hair deflection. Hair shafts are up to 1000 m long. The exact grouping of smooth hairs in rows is typical of the coxae for each pair of legs. 2. Unlike the other, multiply innervated cuticular sensilla of spiders, smooth hairs are supplied by only a single mechanosensitive neuron. This is confirmed by electrophysiological recordings from single hairs. Threshold deflection to elicit a spike response lies near 1°. The response to maintained, step-like stimuli declines rapidly. 3. All central endings of these hair receptors in the fused segmental ganglia are confined to dorsal neuropil of the ipsilateral neuromere. The specific arborization pattern resembles an elongated, three-pronged fork with a long central prong. Topography, natural stimulus situation, and the phasic response characteristic of smooth hairs suggest that spiders use these sensilla to monitor the relative distance between leg coxae during locomotion.     

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.     

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.     

Transplantation of neurons reveals processing areas and rules for synaptic connectivity in the cricket nervous system

In order to assess the nature of spatial cues in determining the characteristic projection sites of sensory neurons in the CNS, we have transplanted sensory neurons of the cricket Acheta domesticus to ectopic locations. Thoracic campaniform sensilla (CS) function as proprioceptors and project to an intermediate layer of neuropil in thoracic ganglia while cercal CS transduce tactile information and project into a ventral layer in the terminal abdominal ganglion (TAG). When transplanted to ectopic locations, these afferents retain their modality-specific projection in the host ganglion and terminate in the layer of neuropil homologous to that of their ganglion of origin. Thus, thoracic CS neurons project to intermediate neuropil when transplanted to the abdomen and cercal CS neurons project to a ventral layer of neuropil when transplanted to the thorax. We conclude that CS can be separated into two classes based on their characteristic axonal projections within each segmental ganglion. We also found that the sensory neurons innervating tactile hairs project to ventral neuropil in any ganglion they encounter after transplantation. Ectopic sensory neurons can form functional synaptic connections with identified interneurons located within the host ganglia. The new contacts formed by these ectopic sensory neurons can be with normal targets, which arborize within the same layer of neuropil in each segmental ganglion, or with novel targets, which lack dendrites in the normal ganglion and are thus normally unavailable for synaptogenesis. These observations suggest that a limited set of molecular markers are utilized for cell–cell recognition in each segmentally homologous ganglion. Regenerating sensory neurons can recognize novel postsynaptic neurons if they have dendrites in the appropriate layer of neuropil. We suggest that spatial constraints produced by the segmentation and the modality-specific layering of the nervous system have a pivotal role in determining synaptic specificity. © 1993 John Wiley & Sons, Inc.