Terminal Abdominal Ganglion Research Articles

Ion transport peptide and ion transport peptide-like regulate ecdysis behavior and water transport during ecdysis in Gryllus bimaculatus

Ion transport peptide (ITP) and ITP-like (ITPLs) are pleiotropic bioactive peptides in insects. Although the contribution of these peptides to ecdysis has been studied, the precise regulatory mechanisms remain poorly understood. Here, we characterized the functions of itp and itpl variants in the two-spotted cricket, Gryllus bimaculatus. Reverse transcription-quantitative PCR and whole-mount in situ hybridization revealed that itp was expressed in the brain and terminal abdominal ganglion, whereas itpl variants were expressed in all ganglia of the central nervous system. Simultaneous knockdown of itp and itpls disrupted ecdysis behavior and water transport from the gut into the hemolymph during molting. Nevertheless, knockdown of itpls without influencing itp expression did not significantly affect ecdysis behavior but caused a reduction in hemolymph mass. Although water transport into the hemolymph is considered necessary for the swelling required to split the old cuticle layers during molting, a rescue experiment by injection of water or cricket Ringer's solution into the hemolymph of knockdown crickets did not recover the normal phenotype. Therefore, we propose that ITP/ITPL control ecdysis behavior probably not by regulating water transport from the gut into the hemolymph in crickets.

Neuromuscular Anatomy and Motor Patterns at the Base of Calling Behaviour in the Female Spongy Moth Lymantria dispar.

"Calling behaviour" is a stereotyped rhythmic motor pattern displayed by female moths, by which they emit the sex pheromone to attract of conspecific males. Calling occurs through a squeezing mechanism based on the turtleneck-like folding and unfolding of the ovipositor cuticle during its telescopic extensions and retractions. This mechanism is under the control of the terminal abdominal ganglion (TAG). By combining anatomical and electrophysiological approaches, here we studied the morpho-functional organisation of the abdominal muscles and the activity of motoneurons from TAG nerve N4-N6 as correlated to the ovipositor movements during calling in the female spongy moth Lymantria dispar. Our results show that the three abdominal segments S7, S8 and S9 (ovipositor) are highly specialized structures containing cuticular appendages, hinges, apodemes and several large muscles, innervated by N4 and especially by N5. N6 mainly innervates the oviductal tract. We also identified a number of motor units from N4 and N5, the spike activity of which is correlated with the ovipositor movements during calling. In conclusion, the release of sex pheromones in the female spongy moth is obtained by extensions and retractions of the ovipositor operated by a coordinated motor program, which is mainly sustained by the activity of a few motor units under the control of TAG nerves N4 and N5.

Functional anatomy of the worker honeybee stinger (Apis mellifera)

The honeybee stinger is a powerful defense mechanism that combines painful venom, a subcutaneous delivery system, and the ability to autotomize. It is a complex organ and to function autonomously it must carry with it all the anatomical components required to operate. In this study, we combined high-speed filming, SEM imagery, and micro-CT for volumetric rendering of the stinger with a synthesis of existing literature. We present a comprehensive description of all components, including cuticular elements, musculature, nervous and glandular tissue using updated imagery. We draw from the Hymenoptera literature to make interspecific comparisons where relevant. The use of 3D reconstruction allows us to separate stinger components and present the first 3D renders of the bee stinger including the terminal abdominal ganglion and its projections. It also clarifies the in-situ geometry of the valves within the bulb and the spatial relationships among the accessory plates and accompanying musculature.

Exploring Honeybee Abdominal Anatomy through Micro-CT and Novel Multi-Staining Approaches.

Simple SummaryApis mellifera or western honeybees are insects belonging to the Order Hymenoptera and the most important pollinators worldwide with great implications in natural biodiversity and agriculture due to their importance in pollination and honey production. The characterization of honeybee anatomy with precise tools will allow a better comprehension of the physiology of these insects under different biological conditions. Here, we employed Micro-computed tomography and novel staining methods to define the morphoanatomical characteristics of the worker honeybee abdomen. We defined the 3D and 2Ds structures of the midgut and hindgut and discovered a new cell type called ventricular telocyte, with possible roles in honeybee epithelium maintenance. Overall, we propose that this method will be useful for further investigation of the structure of the honeybee abdomen under a wide variety of environmental conditions.Continuous improvements in morphological and histochemical analyses of Apis mellifera could improve our understanding of the anatomy and physiology of these insects at both the cellular and tissue level. In this work, two different approaches have been performed to add new data on the abdomen of worker bees: (i) Micro-computed tomography (Micro-CT), which allows the identification of small-scale structures (micrometers) with adequate/optimal resolution and avoids sample damage and, (ii) histochemical multi-staining with Periodic Acid-Schiff-Alcian blue, Lactophenol-Saphranin O and pentachrome staining to precisely characterize the histological structures of the midgut and hindgut. Micro-CT allowed high-resolution imaging of anatomical structures of the honeybee abdomen with particular emphasis on the proventriculus and pyloric valves, as well as the connection of the sting apparatus with the terminal abdominal ganglia. Furthermore, the histochemical analyses have allowed for the first-time description of ventricular telocytes in honeybees, a cell type located underneath the midgut epithelium characterized by thin and long cytoplasmic projections called telopodes. Overall, the analysis of these images could help the detailed anatomical description of the cryptic structures of honeybees and also the characterization of changes due to abiotic or biotic stress conditions.

Central projections of cercal giant interneurons in the adult field cricket, Gryllus bimaculatus.

The structures of neurons, such as dendrites and axonal projections, are closely related to their response properties and their specific functions in neural circuits. Identified neurons, having genetically determined morphological features and pre- and postsynaptic partners, play significant roles in specific behaviors. Giant interneurons (GIs) are identified in the terminal abdominal ganglion of the cricket as mechanosensory projection neurons and are sensitive to airflow stimulation of the cerci. GIs are classified into ventral GIs (vGIs) or dorsal GIs (dGIs) depending on the location of their axons running within the connective nerve cord. Based on their response properties to airflow, vGIs are presumed to be involved in triggering the wind-elicited escape response, whereas dGIs are thought to be airflow direction-encoding neurons. The previous findings regarding airflow sensitivity point to possible differences in the morphology of the central projections that may correspond to their neural functions. However, the detailed morphologies of the GIs in the cephalic and thoracic ganglia of adult crickets remain unclear. In this study, we stained six GIs, namely, GI 8-1 (medial giant interneuron, MGI), 9-1 (lateral giant interneuron, LGI), 9-2, 9-3, 10-2, and 10-3, using intracellular iontophoretic or pressure injection of dyes. Staining revealed remarkable differences in the axonal branching patterns between vGIs and dGIs. The dGIs were further divided into subgroups based on the profiles of their axon collaterals and projection sites in the brain. The anatomical differences between the GIs' central projections seemed to be related to their information encodement and behavioral functions.

Encoding of small-scale air motion dynamics in the cricket, Acheta domesticus.

The cercal sensory system of cricket mediates the detection, localization, and identification of air current signals generated by predators, mates, and competitors. This mechanosensory system has been used extensively for experimental and theoretical studies of sensory coding at the cellular and system levels. It is currently thought that sensory interneurons (INs) in the terminal abdominal ganglion extract information about the direction, velocity, and acceleration of the air currents in the animal's immediate environment and project a coarse-coded representation of those parameters to higher centers. All feature detection is thought to be carried out in higher ganglia by more complex, specialized circuits. We present results that force a substantial revision of current hypotheses. Using multiple extracellular recordings and a special sensory stimulation device, we demonstrate that four well-studied interneurons in this system respond with high sensitivity and selectivity to complex dynamic multidirectional features of air currents that have a spatial scale smaller than the physical dimensions of the cerci. The INs showed much greater sensitivity for these features than for unidirectional bulk-flow stimuli used in previous studies. Thus, in addition to participating in the ensemble encoding of bulk airflow stimulus characteristics, these interneurons are capable of operating as feature detectors for naturalistic stimuli. In this sense, these interneurons are encoding and transmitting information about different aspects of their stimulus environment; they are multiplexing information. Major aspects of the stimulus-response specificity of these interneurons can be understood from the dendritic anatomy and connectivity with the sensory afferent map.NEW & NOTEWORTHY A set of sensory interneurons that have been studied for over 30 years by several different research groups were discovered to have previously unknown encoding characteristics. As well as encoding the direction of bulk airflow with a coarse-coding scheme as shown in previous studies, these interneurons are also responsive to very small-scale, directionally complex air current waveforms. This feature sensitivity can be understood in terms of the cells' complex dendritic branching patterns.

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

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

Distribution of Pheromone Biosynthesis-Activating Neuropeptide in the Central Nervous System of Plutella xylostella (Lepidoptera: Plutellidae).

Insect neuropeptides in the pyrokinin/pheromone biosynthesis-activating neuropeptide (PBAN) family are actively involved in many essential endocrinal functions and serve as potential targets in the search for novel insect control agents. Here, we dissect the nervous system of larval, pupal, and adult Plutella xylostella (L.) (Lepidoptera: Plutellidae) and describe the ganglion morphology and localization of PBAN during different insect developmental stages. Our results show that the central nervous system (CNS) of this species consists of four types of ganglia: cerebral ganglia (brain), subesophageal ganglion (SEG), thoracic ganglia, and abdominal ganglia. A two-lobed brain is connected to the reniform SEG with a nerve cord in larvae and prepupae, whereas in the late pupae and adults, the brain and SEG are fused, forming a brain-SEG complex. The larvae and prepupae have eight abdominal ganglia each, whereas the late pupae and adults each have four abdominal ganglia. Furthermore, all life stages of P. xylostella had similar patterns of PBAN immunoreactivity in the CNS, and the accumulation of PBAN was similar during all life stages except in adult males. PBAN immunoreactive signals were observed in the brain and SEG, and fluorescence signals originating in the SEG extended the entire length of the ventral nerve cord, ending in the terminal abdominal ganglia. Our results provide morphological data that inform the development and evolution of the CNS. In addition, they indicate that the nervous system contains PBAN, which could be used to control P. xylostella populations.

Multiple neuropeptides produced by sex-specific neurons control activity of the male accessory glands and gonoducts in the silkworm Bombyx mori

The male accessory glands (AG) and gonoducts of moths develop during metamorphosis and are essential for successful fertilization of females. We found that these reproductive organs are innervated by a sex-specific cluster of peptidergic neurons in the posterior 9th neuromere of the terminal abdominal ganglion (TAG). This cluster of ~20 neurons differentiate during metamorphosis to innervate the accessory glands and sperm ducts. Using immunohistochemistry and in situ hybridization (ISH) we showed that these neurons express four neuropeptide precursors encoding calcitonin-like diuretic hormone (CT-DH), allatotropin (AT) and AT-like peptides (ATLI-III), allatostatin C (AST-C), and myoinhibitory peptides (MIPs). We used contraction bioassay in vitro to determine roles of these neuropeptides in the gonoduct and accessory gland activity. Spontaneous contractions of the seminal vesicle and AG were stimulated in a dose depended manner by CT-DH and AT, whereas AST-C and MIP elicited dose dependent inhibition. Using quantitative RT-PCR we confirmed expression of receptors for these neuropeptides in organs innervated by the male specific cluster of neurons. Our results suggest a role of these neuropeptides in regulation of seminal fluid movements during copulation.

Defecation initiates walking in the cricket Gryllus bimaculatus

Feces provides information about the donor and potentially attracts both conspecifics and predators and also parasites. The excretory system must be coordinated with other behaviors in insects. We found that crickets started walking forward following defecation. Most intact crickets walked around the experimental arena, stopped at a particular site and raised their bodies up with a slight backward drift to defecate. After the feces dropped to the floor, a cricket started walking with a non-coordinated gait pattern away from the defecation site, and then changed to a tripod gait. To demonstrate that walking is a reflex response to defecation we analyzed the behavior of headless crickets and found that they also showed walking following defecation. In more than half of defecation events, headless crickets walked backwards before defecation. The posture adopted during defecation was similar to that of intact crickets, and forward walking after defecation was also observed. The frequency of forward walking after defecation in headless crickets was greater than in intact crickets. The gait pattern during forward walking was not coordinated and never transitioned to a tripod gait in headless crickets. In animals whose abdominal nerve cords were cut, in any position, pre- or post-defecation walking was not shown in either intact or headless crickets, although they defecated. These results indicated that the terminal abdominal ganglion receives information regarding hind gut condition. It also indicated that ascending signals from the terminal abdominal ganglion initiated leg movement through the neuronal circuits within the thoracic ganglia, and that descending signals from the brain must regulate the leg motor circuit to express the appropriate walking gait.

Octopamine modulates the activity of motoneurons related to calling behavior in the gypsy moth Lymantria dispar.

A morphofunctional investigation of the different neuronal subpopulations projecting through each of the nerves IV-VI emerging bilaterally from the terminal abdominal ganglion (TAG) was correlated with the octopaminergic activity in the ganglion that controls the ovipositor movements associated with calling behavior in the female gypsy moth Lymantria dispar. Tetramethylrodamine-dextran backfills from nerve stumps resulted in a relatively low number of TAG projections, ranging from 12 to 13 for nerve pair IV, 12 to 14 for nerve pair V, and 8 to 9 for nerve pair VI. Furthermore, as assessed by electrophysiological recordings, a number of fibers within each of these nerves displays spontaneous tonic activity, also when the ganglion is fully disconnected from the ventral nerve cord (VNC). Octopamine (OA) applications to the TAG strongly enhanced the activity of these nerves, either by increasing the firing rate of a number of spontaneously firing units or by recruiting new ones. This octopaminergic activity affected calling behavior, and specifically the muscle activity leading to cycling extensions of the intersegmental membrane (IM) between segments VIII and IX (ovipositor). Our results indicate that in the female gypsy moth the octopaminergic neural activity of the TAG is coupled with extensions and retractions of IM for the purpose of releasing pheromone, where motor units innervated by nerve pair IV appear antagonistic with respect to those innervated by nerve pair V.

Oviposition-like central pattern generators in pregenital segments of male and female grasshoppers.

Grasshoppers produce an extraordinary oviposition behavior that is associated with multiple specializations of the skeletal and neuromuscular systems in the posterior abdomen, including a central pattern generator (CPG) in the female's terminal abdominal ganglion. Two pairs of shovel-shaped appendages, the ovipositor valves on the abdomen tip, excavate the soil for deposition of eggs. By contrast, the sexually monomorphic pregenital region of the abdomen is without appendages. Morphological homologues of ovipositor muscles and efferent neurons in the eighth abdominal segment are nevertheless present in pregenital segments of males and females. In both sexes, a robust rhythmic motor program was induced in pregenital segments by the same experimental methods used to elicit oviposition digging. The activity, recorded extracellularly, was oviposition-like in burst period (5-6s) and homologous muscle phase relationships, and it persisted after sensory inputs were removed, indicating the presence of pregenital CPGs. The abdomen exhibited posterior-going waves of activity with an intersegmental phase delay of approximately 1s. These results indicate that serially homologous motor systems, including functional CPGs, provided the foundation for the evolution of oviposition behavior.

Caste differences in the association between dopamine and reproduction in the bumble bee Bombus ignitus

A society of bumble bees is primitively eusocial, with an annual life cycle, and can be used as a physiological model of social bees for comparative studies with highly eusocial hymenopterans. We investigated the dynamics of biogenic amine levels in the brain, meso-metathoracic ganglia, terminal abdominal ganglion, and hemolymph in queens 1 day after mating (1DAM), during diapause (Dp), and during colony founding (CF) in the bumble bee, Bombus ignitus. Dopamine levels in the brain of CF queens were significantly lower than in 1DAM and Dp queens, and the levels in the thoracic ganglia and hemolymph in CF queens were lower than in 1DAM queens, but did not differ from other groups in the abdominal ganglion. Octopamine levels in the brains were higher in Dp queens than in 1DAM queens. Serotonin and tyramine levels did not differ between the groups in different compartments of the central nervous system (CNS) that we examined. The dopamine levels in the brains were significantly positively correlated with those in the thoracic ganglia, abdominal ganglion, and hemolymph, suggesting the regulation of dopamine levels among three different compartments of the CNS. In isolated virgin queens, there were no significant correlations between the brain levels of biogenic amines that we examined and the lengths of the largest terminal oocytes, whereas, in isolated workers, the brain dopamine levels were positively correlated with oocyte lengths. These results suggest that dopamine is associated with ovarian activity in reproductive workers, but not in either virgin or mated queens.

Modulation of Low-Voltage-Activated Inward Current Permeable to Sodium and Calcium by DARPP-32 Drives Spontaneous Firing of Insect Octopaminergic Neurosecretory Cells.

Identification of the different intracellular pathways that control phosphorylation/dephosphorylation process of ionic channels represents an exciting alternative approach for studying the ionic mechanisms underlying neuronal pacemaker activity. In the central nervous system of the cockroach Periplaneta americana, octopaminergic neurons, called dorsal unpaired median (DUM; DUM neurons), generate spontaneous repetitive action potentials. Short-term cultured adult DUM neurons isolated from the terminal abdominal ganglion (TAG) of the nerve cord were used to study the regulation of a tetrodotoxin-sensitive low-voltage-activated (LVA) channel permeable to sodium and calcium (Na/Ca), under whole cell voltage- and current-clamp conditions. A bell-shaped curve illustrating the regulation of the amplitude of the maintained current vs. [ATP]i was observed. This suggested the existence of phosphorylation mechanisms. The protein kinase A (PKA) inhibitor, H89 and elevating [cyclic adenosine 3′, 5′ monophosphate, cAMP]i, increased and decreased the current amplitude, respectively. This indicated a regulation of the current via a cAMP/PKA cascade. Furthermore, intracellular application of PP2B inhibitors, cyclosporine A, FK506 and PP1/2A inhibitor, okadaic acid decreased the current amplitude. From these results and because octopamine (OA) regulates DUM neuron electrical activity via an elevation of [cAMP]i, we wanted to know if, like in vertebrate dopaminergic neurons, OA receptor (OAR) stimulation could indirectly affect the current via PKA-mediated phosphorylation of Dopamine- and cAMP-regulated Phosphoprotein-32 (DARPP-32) known to inhibit PP1/2A. Experiments were performed using intracellular application of phospho-DARPP-32 and non-phospho-DARPP-32. Phospho-DARPP-32 strongly reduced the current amplitude whereas non-phospho-DARPP-32 did not affect the current. All together, these results confirm that DARPP-32-mediated inhibition of PP1/2A regulates the maintained sodium/calcium current, which contributes to the development of the pre-depolarizing phase of the DUM neuron pacemaker activity.

Developmental and sex-specific differences in expression of neuropeptides derived from allatotropin gene in the silkmoth Bombyx mori.

Allatotropin (AT) and related neuropeptides are widespread bioactive molecules that regulate development, food intake and muscle contractions in insects and other invertebrates. In moths, alternative splicing of the at gene generates three mRNA precursors encoding AT with different combinations of three structurally similar AT-like peptides (ATLI-III). We used in situ hybridization and immunohistochemistry to map the differential expression of these transcripts during the postembryonic development of Bombyx mori. Transcript encoding AT alone was expressed in numerous neurons of the central nervous system and frontal ganglion, whereas transcripts encoding AT with ATLs were produced by smaller specific subgroups of neurons in larval stages. Metamorphosis was associated with considerable developmental changes and sex-specific differences in the expression of all transcripts. The most notable was the appearance of AT/ATL transcripts (1) in the brain lateral neurosecretory cells producing prothoracicotropic hormone; (2) in the male-specific cluster of about 20 neurons in the posterior region of the terminal abdominal ganglion; (3) in the female-specific medial neurons in the abdominal ganglia AG2-7. Immunohistochemical staining showed that these neurons produced a mixture of various neuropeptides and innervated diverse peripheral organs. Our data suggest that AT/ATL neuropeptides are involved in multiple stage- and sex-specific functions during the development of B. mori.

Genital Autocleaning in the Male Cricket Gryllus bimaculatus (2): Rhythmic Movements of the Genitalia and Their Motor Control.

Three types of genital movement, their neural controls, and functional roles were investigated to gain a better understanding of the mechanism underlying autocleaning in the male cricket. The membrane complex consisting of the median pouch and genital chamber floor shows peculiar undulation that is composed of two types of movements: a right-left large shift and small crease-like movements. The large shift was caused by contraction of a pair of muscles (MPA) located anterior to the median pouch, while the crease-like movements were caused by numerous muscle fibers extending over the membrane complex. The MPA and muscle fibers were each innervated by efferent neurons in the terminal abdominal ganglion. Experiments with artificial dirt mimicking a foreign object revealed that the crease-like movements were responsible for dirt transport, while the large shift participated in sweeping the dirt into the lateral pouch as a trash container. On the other hand, the dorsal pouch serving as a template for the spermatophore showed a jerky bending movement. Simultaneous monitoring of the membrane complex and dorsal pouch activities suggested that their movements cooperate to enable the efficient evacuation of waste in the dorsal pouch. Based on the results, we conclude that genital autocleaning supports the production of the spermatophore.

Expression of RYamide in the nervous and endocrine system of Bombyx mori

RYamides are neuropeptides encoded by a gene whose precise expression and function have not yet been determined. We identified the RYamide gene transcript (fmgV1g15f, SilkBase database) and predicted two candidates for G-protein coupled RYamide receptors (A19-BAG68418 and A22-BAG68421) in the silkworm Bombyx mori. We cloned the RYamide transcript and described its spatial expression using in situ hybridisation. In the larval central nervous system (CNS) expression of RYamide was restricted to 12–14 small neurons in the brain and two posterior neurons in the terminal abdominal ganglion. During metamorphosis their number decreased to eight protocerebral neurons in the adults. Multiple staining, using various insect neuropeptide antibodies, revealed that neurons expressing RYamide are different from other peptidergic cells in the CNS. We also found RYamide expression in the enteroendocrine cells (EC) of the anterior midgut of larvae, pupae and adults. Two minor subpopulations of these EC were also immunoreactive to antibodies against tachykinin and myosupressin. This expression pattern suggests RYamides may play a role in the regulation of feeding and digestion.

Deep-tissue confocal imaging of the central projections of ovipositor sensory afferents in the Egyptian cotton leafworm, Spodoptera littoralis

The pre-ovipositon behavior of moths is largely dependent upon the cues that a gravid female perceives while assessing potential oviposition sites. Assessment of such sites is accomplished, at least in part, by mechanosensory and gustatory sensilla located on the ovipositor whose sensory neurons project into the terminal abdominal ganglion (TAG). Using anterograde backfill staining, confocal laser scanning microscopy, and three dimensional reconstruction, we traced and analyzed the central projections of the sensory neurons housed in the sensilla located on the ovipositor papillae and explored the neuropilar composition of the TAG in the Egyptian cotton leafworm, Spodoptera littoralis. The TAG consists of three fused neuromeres (6–8th Ner) associated with the 6–8th abdominal segments. Within the TAG, and specifically in the 8th neuromere, four unstructured neuropilar compartments are present; the dorso-ipsilateral motor neuropil (MN), the medio-ipsilateral mechanosensory neuropil (MchN), the medio-ipsilateral small gustatory neuropil (GN), and the medio-contralateral posterior ovipositor glomerulus (Og). The Og appears quite compact, with a hollow core free of terminal arborizations. The MchN is further subdivided into 4 unstructured glomeruli in the 8th neuromere, whose afferents are subsequently extended into 3 glomeruli in the 7th and 6th neuromeres. Few neurites of the Og are populated with large dense varicosities reminiscent of neurosecretory vesicles. Given that all ovipositor nerves converge into a common ganglionic center, the TAG, we assume that this ganglion may be a center for coordination of oviposition behaviors, including movements of the ovipositor during assessment of oviposition substrates and egg laying in S. littoralis.

Exposure to 50Hz electromagnetic field changes the efficiency of the scorpion alpha toxin.

BackgroundExtremely low-frequency (50 Hz) electromagnetic field (ELF-EMF) is produced by electric power transmission lines and electronic devices of everyday use. Some phenomena are proposed as “first effects” of ELF-EMF: the discrete changes in the membrane potential and the increase of the calcium channel activity as well as the intracellular concentration of Ca2+. Interaction of the scorpion alpha toxin with the sodium channel depends on the orientation of the charges and may be perturbed by changes in the membrane polarization. The toxin induces overexcitability in the nervous system and an increase in the neurotransmitters released with different consequences, mainly the paralysis of muscles. We assumed that the exposure to ELF-EMF 0.7 mT will change the effects of the insect selective scorpion alpha toxin (recombinant LqhαIT from Leiurus quinquestriatus hebraeus) at the level of the cercal nerve function, the synaptic transmission and on the level of entire insect organism. Taking into account the compensatory mechanisms in organisms, we tested in addition ten times higher ELF-EMF on whole insects.MethodsExperiments were performed in vivo on cockroaches (Periplaneta americana) and in vitro – on isolated cockroach abdominal nerve cord with cerci. In biotests, the effects of LqhαIT (10−8 M) were estimated on the basis of the insect ability to turn back from dorsal to ventral side. Three groups were compared: the control one and the two exposed to ELF-EMF – 0.7 and 7 mT. Bioelectrical activity of the cercal nerve and of the connective nerve that leaves the terminal abdominal ganglion was recorded using extracellular electrodes. LqhαIT (5 × 10−8 M) induced modifications of neuronal activity that were observed in the control cockroach preparations and in the ones exposed to ELF-EMF (0.7 mT). The exposure to ELF-EMF was carried out using coils with a size appropriate to the examined objects.ResultsThe exposure to ELF-EMF (0.7 mT) modified the effects of LqhαIT (5 × 10−8 M) on activity of the cercal nerve and of the connective nerve. We observed a decrease of the toxin effect on the cercal nerve activity, but the toxic effect of LqhαIT on the connective nerve was increased. Biotests showed that toxicity of LqhαIT (10−8 M) on cockroaches was reduced by the exposure to ELF-EMF (0.7 and 7 mT).ConclusionsThe exposure to 50 Hz ELF-EMF modified the mode of action of the anti-insect scorpion alpha toxin LqhαIT at cellular level of the cockroach nervous system and in biotests. Toxin appeared as a usefull tool in distinguishing between the primary and the secondary effects of ELF-EMF.

Excitatory connections of nonspiking interneurones in the terminal abdominal ganglion of the crayfish.

The output effects of the nonspiking interneurones in the crayfish terminal abdominal ganglion upon the uropod motor neurones were characterized using simultaneous intracellular recordings. Inhibitory interactions from nonspiking interneurones to the uropod motor neurones were one-way and chemically mediated. The depolarization of the motor neurones with current injection increased the amplitude of the nonspiking interneurone-mediated hyperpolarization, while hyperpolarization of the motor neurone decreased it. By contrast, excitatory interactions from the nonspiking interneurones to the motor neurones were not mediated via chemical synaptic transmissions. These excitatory connections with the slow motor neurones were one-way while connections with fast motor neurones were bidirectional. Nonspiking interneurone-mediated membrane depolarization of the motor neurones was not affected by the passage of hyperpolarizing current. Each motor neurone spike elicited a time-locked EPSP in the nonspiking interneurones with very short delay (0.2ms) that suggested electrical coupling between nonspiking interneurones and motor neurones. Nonspiking interneurones directly control the organization of slow motor neurone activity, while they appear to regulate the background activity of the fast motor neurones. A single nonspiking interneurone is possible to inhibit some inter and/or motor neurones via direct chemical synapses and simultaneously excite other neurones via electrical synapses.