Projection areas and branching patterns of the tympanal receptor cells in migratory locusts,Locusta migratoria and Schistocerca gregaria

Summary In Locusta migratoria and Schistocerca gregaria, the projection areas and branching patterns of the tympanal receptor cells in the thoracic ganglia were revealed. Four auditory neuropiles can be distinguished on each side of the ventral cord, always located in the anterior part of the ring tract in each neuromere (two in the meta-, one in the meso-, and one in the prothoracic ganglion). Some of the receptor fibres ascend to the suboesophageal ganglion. There are distinct subdivisions within the auditory, frontal metathoracic and mesothoracic neuropiles. The arrangement of the terminal arborisations of the four types of tympanal receptor cells according to their different frequency-intensity responses is somatotopic and similar in the two ganglia. Here the receptor cells of type-1 form a restricted lateroventral arborisation. Cells of type-4 occupy the caudal part with a dorsorostral extension. Cells of type-2 and -3 arborise in a subdivision between both. Most of the stained low-frequency receptors (type-1, -2, and -3) terminate either in the metathoracic or, predominantly, in the mesothoracic ganglion. In contrast, the high-frequency cells (type-4) ascend to the prothoracic ganglion. The receptor fibres of the different types of receptor cells differ in diameter.Abbreviations aRT anterior part of the ring tract - cf characteristic frequency - MVT median ventral tract - SEG suboesophageal ganglion - SMC supramedian commissure - VMT ventral median tract - VIT ventral intermediate tract Supported by the Deutsche Forschungsgemeinschaft; part of program A7 in Sonderforschungsbereich 305 (Ecophysiology)     

Tympana,auditory thresholds,and projection areas of tympanal nerves in singing and silent grasshoppers (Insecta,Acridoidea)

Summary The auditory systems of several species of singing and acoustically communicating grasshoppers, as well as of silent grasshoppers, were compared with respect to the external structure of the tympana, thresholds of the tympanal nerve response and projection areas of tympanal nerves within the metathoracic part of the ventral nerve cord. Extracellular recordings from the tympanal nerves, using suction electrodes, revealed that singing and silent grasshoppers hear within the frequency range tested, from 2 to 40 kHz. However, differences in sensitivity were observed in those silent species with tympana of modified structure. Cobalt-backfills of the tympanal nerves revealed a clearly discernible auditory neuropil in the anterior ring tract of the metathoracic ganglion in all animals. A comparison of the volumes of neuropilar areas calculated from serial sections of the entire ganglion showed a gradation: the volumes were biggest in singing species, slightly smaller in silent species with a well-developed tympanum, and smallest in the species with modified tympana. These findings support several authors who suggested that auditory organs evolved earlier than acoustic communication.     

Differential Roles for EphA and EphB Signaling in Segregation and Patterning of Central Vestibulocochlear Nerve Projections

Auditory and vestibular afferents enter the brainstem through the VIIIth cranial nerve and find targets in distinct brain regions. We previously reported that the axon guidance molecules EphA4 and EphB2 have largely complementary expression patterns in the developing avian VIIIth nerve. Here, we tested whether inhibition of Eph signaling alters central targeting of VIIIth nerve axons. We first identified the central compartments through which auditory and vestibular axons travel. We then manipulated Eph-ephrin signaling using pharmacological inhibition of Eph receptors and in ovo electroporation to misexpress EphA4 and EphB2. Anterograde labeling of auditory afferents showed that inhibition of Eph signaling did not misroute axons to non-auditory target regions. Similarly, we did not find vestibular axons within auditory projection regions. However, we found that pharmacologic inhibition of Eph receptors reduced the volume of the vestibular projection compartment. Inhibition of EphB signaling alone did not affect auditory or vestibular central projection volumes, but it significantly increased the area of the auditory sensory epithelium. Misexpression of EphA4 and EphB2 in VIIIth nerve axons resulted in a significant shift of dorsoventral spacing between the axon tracts, suggesting a cell-autonomous role for the partitioning of projection areas along this axis. Cochlear ganglion volumes did not differ among treatment groups, indicating the changes seen were not due to a gain or loss of cochlear ganglion cells. These results suggest that Eph-ephrin signaling does not specify auditory versus vestibular targets but rather contributes to formation of boundaries for patterning of inner ear projections in the hindbrain.     

Analysis of ephrin-A2 in the chick retinotectal projection using a function-blocking monoclonal antibody

Eph receptor tyrosine kinases and their ligands have been shown to be involved in processes of cell migration and axon guidance during embryonic development. Here we describe the development of a function-blocking monoclonal antibody against chick ephrin-A2, and its effect on retinal ganglion cell axons studied both in vitro and in vivo. In the stripe assay, the blocking antibody completely abolished the repulsive effect of posterior tectal membranes. In vivo, in a loss-of-function approach, hybridoma cells secreting the antiephrin-A2 antibody were applied to chick embryos from embryonic day 3 (E3) on, and the retinotectal projection was subsequently analyzed at E16. DiI tracing analyses showed that although the projection of both temporal and nasal retinal ganglion axons in the tectum was, overall, normal, occasionally diffuse and extra termination zones were observed, in addition to axons over-shooting their termination zones. These data support the idea that ephrin-A2 contributes to the establishment of the chick retinotectal projection.     

Neuronal connections between central and enteric nervous system in the locust, Locusta migratoria

The number and location of neurons, in the central nervous system, that project into the frontal connective was studied in the locust by using retrograde neurobiotin staining. Staining one frontal connective revealed some 70 neurons in the brain. Most of these were located within both tritocerebral lobes. Additional groups of neurons were located within the deutocerebrum and protocerebrum. Some 60 neurons were labelled in the suboesophageal ganglion. These formed nine discernable populations. In addition, two neurons were located in the prothoracic ganglion and two neurons in the first abdominal neuromere of the metathoracic ganglion. Thus, some 250 neurons located within the head ganglia, and even neurons in thoracic ganglia, project into the ganglia of the enteric nervous system. This indicates that the coordination between the central and enteric ganglia is much more complex than previously thought. With the exception of some previously described dorsal unpaired median neurons and a few motor neurons in the head ganglia, the identity and function of most of these neurons is as yet unknown. Possible functions of the neurons in the thoracic ganglia are discussed.     

Presynaptic contributions to response shape in an auditory neuron of the grasshopper

Neuron 714 is morphologically one of the most prominent neurons in the central auditory pathway of the grasshopper with arborizations extending from the abdominal neuromeres of the metathoracic ganglion to the brain. The aim of this study is to explore auditory information flow involving neuron 714 at the level of the ventral nerve cord. Paired intracellular recordings were made from neuron 714 in the mesothorax on the one hand, and from candidate presynaptic auditory neurons of the metathorax on the other. Electrical stimulation of the tympanal nerves provides an estimate of the synaptic distance between these interneurons and auditory afferents. Four, including neuron 714, are monosynaptically connected to afferents, the remainder disynaptically. Current-injection and spike-triggered averaging reveal that of nine neurons examined, seven make either monosynaptic, disynaptic or polysynaptic connections onto neuron 714. All connections are excitatory. Paired recordings show that response duration and response amplitude in synaptically linked cells vary according to the frequency of the stimulus. Measurements of the latency of the first excitatory post-synaptic potential evoked in neuron 714 by afferents and by metathoracic interneurons show how the synaptic drive from these sources shapes the auditory response of neuron 714. Accepted: 14 October 1998     

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.     

Typical ventilatory pattern of the intact locust is produced by the isolated CNS

Ventilatory rhythms of locusts are generated in the central nervous system (CNS). The primary oscillator or central pattern generator (CPG) is located in the metathoracic ganglion. We studied the different patterns of ventilation by recording long-term efferent discharges from the isolated metathoracic ganglion.Two different basic patterns occur: continuous ventilation and discontinuous ventilation. These patterns can be found in the isolated nerve cord as well as in intact animals. In intact animals sensory feedback usually elicits high frequency continuous ventilation as is the case in most physiological experiments. Many studies of ventilation-associated interneurones were performed under what we call stressed conditions i.e. with strong sensory feedback. Under these conditions many interneurones may be recruited which probably do not belong to the basic CPG. In isolated nerve cords of locusts we recognised the two basic types of ventilation. This provides an experimental approach to the origin of rhythmogenesis in ventilation. We can now examine single interneurones under less stressed or even discontinuous ventilatory conditions in the isolated CNS.We suggest the dominance of intrinsic rhythmogenesis of ventilation in the metathoracic ganglion of locusts.     

The midline metathoracic ear of the praying mantis,Mantis religiosa

Summary The praying mantis, Mantis religiosa, is unique in possessing a single, tympanal auditory organ located in the ventral midline of its body between the metathoracic coxae. The ear is in a deep groove and consists of two tympana facing each other and backed by large air sacs. Neural transduction takes place in a structure at the anterior end of the groove. This tympanal organ contains 32 chordotonal sensilla organized into three groups, two of which are 180° out of line with the one attaching directly to the tympanum. Innervation is provided by Nerve root 7 from the metathoracic ganglion. Cobalt backfills show that the auditory neuropile is a series of finger-like projections terminating ipsilaterally near the midline, primarily near DC III and SMC. The auditory neuropile thus differs from the pattern common to all other insects previously studied.     

Structure of cephalic ganglia and their changes in the post-embryonic development of <Emphasis Type="Italic">Calliphora vomitoria</Emphasis> (L.) (Diptera,Calliphoridae)

The central nervous system of Calliphora vomitoria larvae is situated in the metathoracic and the first abdominal segments and is characterized by a high degree of oligomerization. It consists of only two ganglia: the supraoesophageal ganglion, or brain, and one large synganglion, a product of fusion of the suboesophageal ganglion, three thoracic, and all the abdominal ganglia. Weak development of the neuropil of the larval optic and olfactory lobes in the supraoesophageal ganglion is the result of a significant reduction of the head capsule and sensory organs in the larvae. The formation of the imaginal optic lobes begins at the III larval instar. The commissure of the future central body is present in the I instar larva, but formation of the imaginal structure of the central complex proceeds in the 3-day pupae and ends at the late pupal stage. The mushroom bodies are represented in the I instar larvae only by the pedunculi; the calyces can be distinguished in the II instar larvae but the final formation of their structure and the lobes of the imaginal type occurs at the pupal stage. The glomeruli in the deutocerebrum are also formed at a late stage of pupal development. Based on the degree of development of ganglia of the central nervous system, we can conclude that individual development of higher Diptera is characterized by deep de-embryonization.     

NMDA receptor antagonists disrupt the retinotectal topographic map

We tested the effect of two NMDA receptor antagonists, APV or MK801 (with NMDA), and the receptor agonist NMDA on the maintenance of retinal topography in frogs. Topography was assayed by measuring the dispersion of retrogradely labeled ganglion cells following a local HRP injection into the tectum. In untreated tadpoles, labeled cells covered about 5% of the retinal area. In APV- or MK801/NMDA-treated tadpoles, labeled ganglion cells covered 17% and 10% of the retinal area, respectively. Neither treatment with L-APV nor with NMDA disrupts the fidelity of the retinotectal projection. Neither APV- nor NMDA-treated ganglion cell terminals differed from untreated terminals with respect to tangential area, branch number, or branch density. These data support a role for the NDMA receptor in visual system development.     

Analysis of ephrin‐A2 in the chick retinotectal projection using a function‐blocking monoclonal antibody

Eph receptor tyrosine kinases and their ligands have been shown to be involved in processes of cell migration and axon guidance during embryonic development. Here we describe the development of a function‐blocking monoclonal antibody against chick ephrin‐A2, and its effect on retinal ganglion cell axons studied both in vitro and in vivo. In the stripe assay, the blocking antibody completely abolished the repulsive effect of posterior tectal membranes. In vivo, in a loss‐of‐function approach, hybridoma cells secreting the antiephrin‐A2 antibody were applied to chick embryos from embryonic day 3 (E3) on, and the retinotectal projection was subsequently analyzed at E16. DiI tracing analyses showed that although the projection of both temporal and nasal retinal ganglion axons in the tectum was, overall, normal, occasionally diffuse and extra termination zones were observed, in addition to axons over‐shooting their termination zones. These data support the idea that ephrin‐A2 contributes to the establishment of the chick retinotectal projection. © 2001 John Wiley & Sons, Inc. J Neurobiol 47: 245–254, 2001     

Phylogenetic,ontogenetic and adult adaptive plasticity of rhythmic neural networks: a common neuromodulatory mechanism?

Neuromodulatory inputs are known to play a major role in the adaptive plasticity of rhythmic neural networks in adult animals. Using the crustacean stomatogastric nervous system, we have investigated the role of modulatory inputs in the development of rhythmic neural networks. We found that the same neuronal population is organised into a single network in the embryo, as opposed to the two networks present in the adult. However, these adult networks pre-exist in the embryo and can be unmasked by specific alterations of the neuromodulatory environment. Similarly, adult networks may switch back to the embryonic phenotype by manipulating neuromodulatory inputs. During development, we found that the early established neuromodulatory population display alteration in expressed neurotransmitter phenotypes, and that although the population of modulatory neurones is established early, with morphology and projection pattern similar to adult ones, their neurotransmitter phenotype may appear gradually. Therefore the abrupt switch from embryonic to adult network expression occurring at metamorphosis may be due to network reconfiguration in response to changes in modulatory input, as found in adult adaptive plasticity. Strikingly, related crustacean species express different motor outputs using the same basic network circuitry, due to species-specific alteration in neuromodulatory substances within homologous projecting neurones. Therefore we propose that alterations within neuromodulatory systems to a given rhythmic neural network displaying the same basic circuitry may account for the generation of different motor outputs throughout development (ontogenetic plasticity), adulthood (adaptive plasticity) and evolution (phylogenetic plasticity).Abbreviations CoG Commissural ganglion - OG Oesophageal ganglion - STG Stomatogastric ganglion - STNS Stomatogastric nervous system     

Morphological and physiological regeneration in the auditory system of adult <Emphasis Type="Italic">Mecopoda elongata</Emphasis> (Orthoptera: Tettigoniidae)

Orthopterans are suitable model organisms for investigations of regeneration mechanisms in the auditory system. Regeneration has been described in the auditory systems of locusts (Caelifera) and of crickets (Ensifera). In this study, we comparatively investigate the neural regeneration in the auditory system in the bush cricket Mecopoda elongata. A crushing of the tympanal nerve in the foreleg of M. elongata results in a loss of auditory information transfer. Physiological recordings of the tympanal nerve suggest outgrowing fibers 5 days after crushing. An anatomical regeneration of the fibers within the central nervous system starts 10 days after crushing. The neuronal projection reaches the target area at day 20. Threshold values to low frequency airborne sound remain high after crushing, indicating a lower regeneration capability of this group of fibers. However, within the central target area the low frequency areas are also innervated. Recordings of auditory interneurons show that the regenerating fibers form new functional connections starting at day 20 after crushing.     

The cytokine macrophage migration inhibitory factor (MIF) acts as a neurotrophin in the developing inner ear of the zebrafish, Danio rerio

Macrophage migration inhibitory factor (MIF) plays versatile roles in the immune system. MIF is also widely expressed during embryonic development, particularly in the nervous system, although its roles in neural development are only beginning to be understood. Evidence from frogs, mice and zebrafish suggests that MIF has a major role as a neurotrophin in the early development of sensory systems, including the auditory system. Here we show that the zebrafish mif pathway is required for both sensory hair cell (HC) and sensory neuronal cell survival in the ear, for HC differentiation, semicircular canal formation, statoacoustic ganglion (SAG) development, and lateral line HC differentiation. This is consistent with our findings that MIF is expressed in the developing mammalian and avian auditory systems and promotes mouse and chick SAG neurite outgrowth and neuronal survival, demonstrating key instructional roles for MIF in vertebrate otic development.     

Changes in the auditory neuropil after deafferentation in adult grasshoppers (Schistocerca gregaria)

Nervous systems are capable of structural adjustments. Such plastic changes also occur in the auditory system of the locust Schistocerca gregaria in which a deafferentation leads to compensatory mechanisms, such as collateral sprouting of interneurons. In this study we further investigated lesion related changes in the major auditory neuropil, the median ventral association center (mVAC) of the metathoracic ganglion. The auditory sensory organ of adult locusts was unilaterally extirpated and the mVAC was histologically and immunocytochemically analyzed until 20 days postoperative. Measurements of the neuropil area in transverse sections showed a decrease in size. The putative transmitter of the afferents, acetylcholine, was investigated by acetylcholinesterase histochemistry. Comparisons of staining intensities in the intact and deafferentated mVAC indicated that the amount of acetylcholinesterase in the deafferentated mVAC decreased shortly after the operation. Both, the decreases in size of the mVAC as well as that in acetylcholinesterase histochemistry were only less than 10% compared to the controls. The immunoreactivity against the neurotransmitters γ-amino butyric acid and serotonin was not influenced by the deafferentation.     

Regeneration of the projection and synaptic connections of tympanic receptor fibers of Locusta migratoria (Orthoptera) after axotomy

The tergite nerve N6 of the first abdominal segment of the locust Locusta migratoria contains receptor fibers, from the tympanic organ, and hair sensilla as well as motoric axons. The nerve was axotomized in nymphal instars or adults, and the regeneration of nerve fibers was studied. The sensory fibers regrow and regenerate their projection pattern within the central nervous system. They recognize their specific neuropile areas even after entering the ganglion through different pathways. The receptor fibers of the tympanic organ reestablish synaptic connections to auditory interneurons, even though the physiological characteristics of the interneurons are not fully restored. This regenerative capability contrasts with the lack of regeneration of peripheral structures in locusts, but supports the described plasticity in the auditory system of monaural locusts (Lakes, Kalmring, and Engelhard, 1990). The motor fibers do not regenerate nerves innervating muscles of the body wall.     

Position,guidance, and mapping in the developing visual system

Positional identity in the visual system affects the topographic projection of the retina onto its central targets. In this review we discuss gradients and positional information in the retina, when and how they arise, and their functional significance in development. When the axons of retinal ganglion cells leave the eye, they navigate through territory in the central nervous system that is rich in positional information. We review studies that explore the navigational cues that the growth cones of retinal axons use to orient towards their target and organize themselves as they make this journey. Finally, these axons arrive at their central targets and make a precise topographic map of visual space that is crucial for adaptive visual behavior. In the last section of this review, we examine the topographic cues in the tectum, what they are, when, and how they arise, and how retinal axons respond to them. We also touch on the role of neural activity in the refinement of this topography. © 1993 John Wiley & Sons, Inc.     

Control of melanization in first instar larvae of Schistocerca gregaria

Melanization in first-instar larvae of Schistocerca is controlled by a hormone released from neurosecretory axon terminals of the fine nerves posterior to the metathoracic ganglion. The hormone is not detectable in the haemolymph before the embryonic ecdysis but is present within seconds after the ecdysis has started. It is suggested that horizontal displacement of the embryonic cuticle is the trigger for the release of the hormone and that the prothoracic ganglion forms part of the neural pathway between the sensory input caused by ecdysis and the release of the hormone.