New vistas on the initiation and maintenance of insect motor behaviors revealed by specific lesions of the head ganglia

In insects, thoracic pattern generators are modulated by the two head ganglia, the supraesophageal ganglion (brain) and the subesophageal ganglion, which act as higher-order neuronal centers. To explore the contribution of each head ganglion to the initiation and maintenance of specific motor behaviors in cockroaches (Periplaneta americana), we performed specific lesions to remove descending inputs from either the brain or the subesophageal ganglion or both, and quantified the behavioral outcome with a battery of motor tasks. We show that ‘emergency’ behaviors, such as escape, flight, swimming or righting, are initiated at the thoracic level independently of descending inputs from the head ganglia. Yet, the head ganglia play a major role in maintaining these reflexively initiated behaviors. By separately removing each of the two head ganglia, we show that the brain excites flight behavior and inhibits walking-related behaviors, whereas the subesophageal ganglion exerts the opposite effects. Thus, control over specific motor behaviors in cockroaches is anatomically and functionally compartmentalized. We propose a comprehensive model in which the relative permissive versus inhibitory inputs descending from the two head ganglia, combined with thoracic afferent sensory inputs, select a specific thoracic motor pattern while preventing the others.     

Organizational principles of outputs from Dipteran brains

Two major classes of Descending Neurones (DNs) originate in fly cerebral ganglia: (1) uniquely identifiable DNs, most of which arise preorally in duetocerebral neuropil of the supraoesophageal ganglion, the brain proper (2) parallel projecting DNs (PDNs) most originating in the suboesophageal ganglion. Brain DNs receive inputs directly from sensory systems and indirectly via higher center and peptidergic interconnections of the protocerebrum. Direct inputs include primary mechanosensory afferents, first order relay neurones from the olfactory lobes and ocellar receptor cups, and higher order visual neurones that interact with retinotopic inputs from compound eyes. Uniquely identifiable DNs arising in the brain are arranged in uniquely identifiable clusters. Each cluster receives a unique combination of inputs which are shared wholly or in part by the dendritic trees of its constituent DNs. Axons arising from a cluster diverge to different targets in the thoracic ganglia. PDNs form groups of as many as 40 neurones, as determined from outgoing axon bundles. Dendrites of PDNs are thin and diffuse, and arborize amongst collaterals from through-going axons of descending neurones arising in the brain. Axon bundles of PDNs are typically organized in rather simple ladder-like patterns in thoracic ganglion. A third type of uniquely identifiable DN also arises in the suboesophageal ganglion but does not seem to be arranged in clusters.     

Insects and Vertebrates: Analogous Structures in Higher Integrative Centers of the Brain

This work deals with studies on anatomical relationships, neuronal composition, and some synaptic connections that exist in the central complex (CC) in the supraesophageal ganglion in larva of dragonfly g. Aeschna. It has been shown that CC contains protocerebral bridge of an elongated and slightly curved cylindrical shape, fan-shaped and ellipsoid bodies of a bean-like shape and two small roundish noduli. There were revealed (stained) neurons providing both internal connections of CC and its connections with other CNS regions. Connections with tritocerebrum, the higher center of the autonomic nervous system, and subesophageal ganglion, an intermediate relay between supraesophageal ganglion and truncal brain, have been established. The existence of connections of CC with nuclei of abdominal nervous chain is suggested. Connection of ocelli with the CC has been traced. Unipolar neurons of the same type have been revealed, each of them giving collaterals to protocerebral bridge and ending as bushy terminals that form the main part of glomerule in the fan-shaped and ellipsoid bodies. Glomeruli are arranged in rows, in which cross connections have been found. It has been established that the structure of neuropils of the fan-shaped and ellipsoid bodies represent a shielding structure described in the cerebral cortex, midbrain cortex, and cerebellar cortex of vertebrates. Thus, in insects, like in vertebrates, the shielding structures developed not only in optic centers, but also in structures performing higher integrative functions. A possible functional role of the central complex is discussed.     

Spinal projections of brainstem respiratory related neurons in the cat as revealed by retrograde fluorescent markers

By means of retrograde axonal transport of fluorescent tracers, connections between brainstem respiratory related regions and the spinal cord has been studied in the cat. Neurons at the pneumotaxic center project bilaterally (90% ipsi-, 10% contra-) to cervical and lumbar spinal cord and ipsilaterally to thoracic levels. The ventrolateral nucleus of the tractus solitarius project mainly contralaterally (85%) to cervical levels and only contralaterally to thoracic levels; no efferent projections were found to lumbar levels. The ventral respiratory group showed a great number of neurons projecting to the spinal cord especially from the nucleus retroambiguus. Both nuclei, ambiguus and retroambiguus, project mainly contralaterally (70%) to the spinal cord. The B?tzinger complex showed rather scarce bilateral projections to cervical and only ipsilateral projections to lower cervical, thoracic and lumber levels.     

The identification and distribution of gonadotropin-releasing hormone-like peptides in the central nervous system and ovary of the giant freshwater prawn, Macrobrachium rosenbergii

In the present study, we demonstrated the existence of GnRH-like peptides in the central nervous system (CNS) and ovary of the giant freshwater prawn, Macrobrachium rosenbergii using immunocytochemistry. The immunoreactivity (ir) of lamprey (l) GnRH-III was detected in the soma of medium-sized neurons located in neuronal cluster number 11 in the middle part of supraesophageal ganglion (deutocerebrum), whereas ir-octopus (oct) GnRH was observed in the soma of both medium-sized and large-sized neurons in thoracic ganglia, as well as in the fibers innervating the other medium-sized and large-sized neuronal cell bodies in the thoracic ganglia. In addition, ir-lGnRH-I was observed in the cytoplasm of late previtellogenic oocyte and early vitellogenic oocyte. These data suggest that M. rosenbergii contain at least three isoforms of GnRH: two GnRH isoforms closely related to lGnRH-III and octGnRH in the CNS, whereas another isoform, closely related to lGnRH-I, was localized in the ovary. This finding provides supporting data that ir-GnRH-like peptide(s) may exist in this decapod crustacean.     

A Wasp Manipulates Neuronal Activity in the Sub-Esophageal Ganglion to Decrease the Drive for Walking in Its Cockroach Prey

Background

The parasitoid Jewel Wasp hunts cockroaches to serve as a live food supply for its offspring. The wasp stings the cockroach in the head and delivers a cocktail of neurotoxins directly inside the prey''s cerebral ganglia. Although not paralyzed, the stung cockroach becomes a living yet docile ‘zombie’, incapable of self-initiating spontaneous or evoked walking. We show here that such neuro-chemical manipulation can be attributed to decreased neuronal activity in a small region of the cockroach cerebral nervous system, the sub-esophageal ganglion (SEG). A decrease in descending permissive inputs from this ganglion to thoracic central pattern generators decreases the propensity for walking-related behaviors.

Methodology and Principal Findings

We have used behavioral, neuro-pharmacological and electrophysiological methods to show that: (1) Surgically removing the cockroach SEG prior to wasp stinging prolongs the duration of the sting 5-fold, suggesting that the wasp actively targets the SEG during the stinging sequence; (2) injecting a sodium channel blocker, procaine, into the SEG of non-stung cockroaches reversibly decreases spontaneous and evoked walking, suggesting that the SEG plays an important role in the up-regulation of locomotion; (3) artificial focal injection of crude milked venom into the SEG of non-stung cockroaches decreases spontaneous and evoked walking, as seen with naturally-stung cockroaches; and (4) spontaneous and evoked neuronal spiking activity in the SEG, recorded with an extracellular bipolar microelectrode, is markedly decreased in stung cockroaches versus non-stung controls.

Conclusions and Significance

We have identified the neuronal substrate responsible for the venom-induced manipulation of the cockroach''s drive for walking. Our data strongly support previous findings suggesting a critical and permissive role for the SEG in the regulation of locomotion in insects. By injecting a venom cocktail directly into the SEG, the parasitoid Jewel Wasp selectively manipulates the cockroach''s motivation to initiate walking without interfering with other non-related behaviors.     

Surgical generation of supernumerary appendages for studying neuronal specificity in Drosophila melanogaster

The ability of sensory neurons to establish specific synaptic contacts in the central nervous system (CNS) can be studied by changing the spatial relationship between the periphery and the CNS. In contrast to the genetic displacement of appendages by homoeotic mutations, the surgical approach used in this study allows one to place homologous as well as heterologous appendages to the same site on the body surface. Using an improved technique of “surface transplantation,” we generated supernumerary appendages of any desired type in a particular abdominal position. The sensory axons originating from these grafts enter the CNS through the main abdominal nerve and arborize in the fused abdominal ganglia; many fibers extend also into thoracic centers. In the abdominal ganglia, terminals from dorsal transplants (wings and halteres) stay on the ipsilateral side, whereas terminals from ventral transplants (legs and antennae) distribute ipsi- and contralaterally. The same preference holds true for dorsal and ventral abdominal bristles, respectively, whose projection patterns served as a reference. In thoracic ganglia, axons from dorsal and ventral grafts yield completely different terminal patterns. Dorsal grafts project into the ipsilateral wing center, even in the mutant wingless, in which normal wing afferents are suppressed. In contrast, fibers from ventral grafts often extend along the thoracic midline. These data indicate that sensory axons of homologous appendages on the one hand, and their central targets on the other, share serially repeated surface markers. This may enable sensory fibers to recognize centers of homologous appendages.     

Effects of neck and circumoesophageal connective lesions on posture and locomotion in the cockroach

Few studies in arthropods have documented to what extent local control centers in the thorax can support locomotion in absence of inputs from head ganglia. Posture, walking, and leg motor activity was examined in cockroaches with lesions of neck or circumoesophageal connectives. Early in recovery, cockroaches with neck lesions had hyper-extended postures and did not walk. After recovery, posture was less hyper-extended and animals initiated slow leg movements for multiple cycles. Neck lesioned individuals showed an increase in walking after injection of either octopamine or pilocarpine. The phase of leg movement between segments was reduced in neck lesioned cockroaches from that seen in intact animals, while phases in the same segment remained constant. Neither octopamine nor pilocarpine initiated changes in coordination between segments in neck lesioned individuals. Animals with lesions of the circumoesophageal connectives had postures similar to intact individuals but walked in a tripod gait for extended periods of time. Changes in activity of slow tibial extensor and coxal depressor motor neurons and concomitant changes in leg joint angles were present after the lesions. This suggests that thoracic circuits are sufficient to produce leg movements but coordinated walking with normal motor patterns requires descending input from head ganglia.Electronic Supplementary Material Supplementary material is available for this article at     

Assembly and breakdown of Cajal bodies in accessory nuclei of Hymenoptera

In some species of insects, oocytes have vesicular organelles, termed accessory nuclei (ANs). The ANs form by budding off from the nuclear envelope of the oocyte and are filled with translucent matrix containing dense inclusions. One type of these inclusions contains coilin and small nuclear ribonucleoproteins (snRNPs) and is homologous to Cajal bodies. We describe the early events in the morphogenesis of Cajal bodies in the ANs (ANCBs) of the common wasp, Vespula germanica, and show that they contain survival of motor neurons (SMN) protein. We present evidence that in the wasp, ANCBs form by the gradual accumulation of aggregates composed of SMN and small nuclear RNAs. We also show that ANCBs break down and disperse within the ANs as the ANs, which initially surround the oocyte nucleus, localize to the oocyte cortex. The components of dispersed ANCBs are retained within ANs until the end of oogenesis, which suggests that their function may be required at the onset of embryonic development. Because the morphology and behavior of ANs and their Cajal body-like inclusions are conserved in two other hymenopteran species, these features might be characteristic of all hymenopterans.     

Anatomical and functional characterisation of the stomatogastric nervous system of blowfly (Calliphora vicina) larvae

The anatomy and functionality of the stomatogastric nervous system (SNS) of third-instar larvae of Calliphora vicina was characterised. As in other insects, the Calliphora SNS consists of several peripheral ganglia involved in foregut movement regulation. The frontal ganglion gives rise to the frontal nerve and is connected to the brain via the frontal connectives and antennal nerves (ANs). The recurrent nerve connects the frontal- to the hypocerebral ganglion from which the proventricular nerve runs to the proventricular ganglion. Foregut movements include rhythmic contractions of the cibarial dilator muscles (CDM), wavelike movements of crop and oesophagus and contractions of the proventriculus. Transections of SNS nerves indicate mostly myogenic crop and oesophagus movements and suggest modulatory function of the associated nerves. Neural activity in the ANs, correlating with postsynaptic potentials on the CDM, demonstrates a motor pathway from the brain to CDM. Crop volume is monitored by putative stretch receptors. The respective sensory pathway includes the recurrent nerve and the proventricular nerve. The dorsal organs (DOs) are directly connected to the SNS. Mechanical stimulation of the DOs evokes sensory activity in the AN. This suggests the DOs can provide sensory input for temporal coordination of feeding behaviour.     

The distribution of several amino acids in specific ganglia and nerve bundles of the lobster

Using the technique of measuring DNP-amino acid methyl esters by gas-liquid chromatography, the distribution of alanine, proline, glycine, GABA, glutamate and aspartate was determined in individual ganglia and the associated nerve bundles between these ganglia after isolation from the nervous system of the lobster, Homarus americanus. The brain or supraesophageal ganglion (27.2 mg) and the next 5 thoracic ganglia (varying from 24 to 10 mg in a rostral–caudal direction) as well as the nerve bundles connecting these ganglia were used. GABA and aspartate values varied the most among the individual ganglia; highest values were found in the second and third thoracic ganglia. The levels of alanine, proline, glycine and glutamate varied very little from ganglion to ganglion; however, the values for these amino acids did exhibit some variability among the individual connectives. The highest value for each was in the nerve bundle between the first and second thoracic ganglion. Glycine was present at the highest level of any of the amino acids whereas GABA was at the lowest level in the individual structures assayed.     

Morphological and physiological properties of the giant interneuron of the hermit crab (Pagurus pollicaris)

The physiological and morphological properties of the giant interneurons in the hermit crab Pagurus pollicaris are described. The cell bodies are located anteriorly in the supraesophageal ganglion, close to the mid-line. Each cell sends a neurite posteriorly and then laterally, so that they cross over in the center of the ganglion. Each axon then branches: one branch runs laterally while the other travels posteriorly and leaves the ganglion in the circumesophageal connective on the side contralateral to the cell body. The giant axons travel in the circumesophageal connectives and through the thoracic and abdominal ganglia without branching. Each giant axon makes synaptic contact with its ipsilateral giant abdominal flexor motor neuron and with a second flexor motor neuron that has its axon in the contralateral third root. In the supraesophageal ganglion there is a bidirectional synapse between the two giant interneurons. Intracellular recordings from the giant axons show that there is a delay of 0.5 to 0.75 ms that cannot be accounted for by spike propagation along the axons, and may be accounted for by a chemical synapse between the giant interneurons.     

Afferent projections of infrared-sensitive sensilla in the beetle Melanophila acuminata (Coleoptera: Buprestidae)

Beetles of the genus Melanophila are able to detect infrared radiation by using specialized sensilla in their metathoracic pit organs. We describe the afferent projections of the infrared-sensitive neurons in the central nervous system. The axons primarily terminate in the central neuropil of the fused second thoracic ganglia where they establish putative contacts with ascending interneurons. Only a few collaterals appear to be involved in local (uniganglionic) circuits. About half of the neurons send their axons further anterior to the prothoracic ganglion. A subset of these ascend to the subesophageal ganglion, and about 10% project to the brain. Anatomical similarities suggest that the infrared-sensitive neurons are derived from neurons supplying mechanosensory sensilla. The arborization pattern of the infrared afferents suggests that infrared information is processed and integrated upstream from the thoracic ganglia.     

Statocysts in crabs: short-term control of locomotion and long-term monitoring of hydrostatic pressure

Crabs show well-coordinated locomotion. They have proprioceptors similar to those of lobsters, but they differ in terms of their balancing systems and their condensed nervous system, which allows rapid interganglionic conduction. Typically they exhibit dynamically stable locomotion with a highly developed semicircular canal system that codes angular acceleration in each of three orthogonal planes (horizontal and vertical at 45 degrees and 135 degrees to the pitching plane). Left and right interneurons each code one direction of angular acceleration, carrying information between the brain and the thoracic ganglia. Cell A codes head-up vertical plane angular accelerations. Cell B codes rotations in the horizontal plane. Interneurons C and D code headdown vertical plane information, carrying it ipsilaterally and contralaterally respectively. These interneurons have a central role in locomotion. They are activated and have their responsiveness to angular acceleration enhanced before and during locomotion. Such simple activation pathways point to how an angular-acceleration-controlled robot (CRABOT) could be constructed. Hydrostatic pressure information carried by the thread hairs, which also sense angular acceleration, is filtered out from direct pathways onto the interneurons, but spectral analysis shows that it still has an influence via central pathways. Long-term recordings from equilibrium interneurons in free-walking crabs taken from the wild into constant conditions show tidally changing frequencies     

Origin and clonal relationship of common inhibitory motoneurons CI1 and CI3 in the locust CNS

In the primordial thoracic ganglia of locust embryos, the bromodeoxiuridine (BrdU) technique for labelling proliferating cells and their progeny was combined with intracellular dye injection to investigate the origin and the clonal relationship of common inhibitory motoneurons. Common inhibitors 1 (CI₁) and 3 (CI₃) were found to be siblings, that is, they are produced by the division of one ganglion mother cell. This ganglion mother cell results from the first division of neuroblast 5–5, at about 30% of embryonic development. A large portion, at least, of the ganglion mother cells produced by subsequent divisions of neuroblast 5–5 give rise to interneurons with contralaterally ascending or descending axons and GABA-like immunoreactivity. Thus, CI₁ and CI₃ are more closely related to putative inhibitory interneurons than they are to other, that is, excitatory, motoneurons. Consistent with this, the CI somata are associated with cell bodies of putative inhibitory interneurons rather than with clusters of excitatory motoneuron somata. These results elicit speculations regarding the evolutionary origin of inhibitory motoneurons. 1994 John Wiley & Sons, Inc.     

Primary sensory projections from the labella to the brain of Drosophila melanogaster Meigen (Diptera : Drosophilidae)

Golgi silver impregnation of sensory neurons arising from labellar taste sensilla of Drosophila melanogaster Meigen (Diptera : Drosophilidae) revealed 7 distinct types I-VII of primary (sensory) fibres projecting to the suboesophageal ganglion (SOG) of the brain. Each fibre was classified on the bases of the neuropil volume occupied by its terminal arborisation, the shape of neuropil region receiving the arborisations and the detailed morphology of the arborisations. The primary sensory fibre projections from the labella are confined to the SOG where they project mainly in the anterior and central neuropils. No labellar sensory fibres project to posterior SOG. Of these 7 types of sensory fibres, three (III, IV and VII) show ipsilateral projections, while others have both ipsi-, and contralateral branches.Four types of interneurons are suggested to be associated with taste perception. Type A interneurons are local interneurons with arborisations confined only to the taste sensory neuropil of the SOG. The types B - D interneurons are interganglionic/output neurons with axons projecting to various brain regions-SOG, calyces of the mushroom bodies, tritocerebrum and thoracic ganglia. These projections suggest that more than one centre (SOG, tritocerebrum, calyces of the mushroom bodies and thoracic ganglia) are involved in processing gustatory information.     

Striatal dopamine levels after unilateral lesions of the substantia nigra: evidence for a contralateral decrease

Unilateral electrolytical and chemical (6-hydroxydopamine) lesions in the substantia nigra (SN) of rats were followed 7 days later by considerable bilateral decreases of neostriatal dopamine (DA) levels. Similarly, the DA content of the substantia nigra decreased not only ipsilaterally but contralaterally as well. Positive correlations were found between ipsi- and contralateral nigral DA levels, ipsi- and contralateral striatal DA and between the DA level of the SN and the striatum of the corresponding side both ipsi- and contralaterally to the lesion.     

CNS microstructure in the wandering wolf spider Arctosa kwangreungensis (Araneae: Lycosidae)

The supraesophageal ganglion of the wolf spider Arctosa kwangreungensis is made up of a protocerebral and tritocerebral ganglion, whereas the subesophageal ganglionic mass is composed of a single pair of pedipalpal ganglia, four pairs of appendage ganglia, and a fused mass of abdominal neuromeres. In the supraesophageal ganglion, complex neuropile masses are located in the protocerebrum which include optic ganglia, the mushroom bodies, and the central body. Characteristically, the only nerves arising from the protocerebrum are the optic nerves, and the neuropiles of the principal eyes are the most thick and abundant in this wandering spider. The central body which is recognized as an important association center is isolated at the posterior of the protocerebrum and appears as a complex of highly condensed neurons. These cells give off fine parallel bundles of axons arranged in the mushroom bodies. The subesophageal nerve mass can be divided into two main tracts on the basis of direction of the neuropiles. The dorsal tracts are contributed to from the motor or interneurons of each ganglion, whereas the ventral tracts are from incoming sensory axons.     

FMRfamide-like immunoreactivity in the ventral nerve cord of the larval eastern spruce budworm,Choristoneura fumiferana (clemens) (Lepidoptera : Tortricidae)

Distribution of FMRFamide-like immunoreactivity was examined in the larval ventral nerve cord of the eastern spruce budworm, Choristoneura fumiferana (Lepidoptera : Tortricidae). Indirect immunofluorescent methods revealed the existence of 3 groups of FLI neurons in each ganglion. The neurons are distributed in a bilaterally symmetrical fashion at the anterodorsal, midlateral and posteroventral regions of the ganglia. There are 4 FMRFamide-like immunoreactive fiber tracts on the dorsal surface of the ganglia to which the anterodorsal FLI neurons project ipsilaterally, while the midlateral pair projects both ipsi-, and contralaterally. The last abdominal ganglion (AG8) has 4 additional pairs of FLI neurons; and axons from some of these extend into the median abdominal nerve, which suggests some role for this neuropeptide in the control of posterior structures of the larva.