Abdominal Ganglion Research Articles

Analysis of temperature-dependent abnormal bursting patterns of neurons in Aplysia.

Temperature affects the firing pattern and electrical activity of neurons in animals, eliciting diverse responses depending on neuronal cell type. However, the mechanisms underlying such diverse responses are not well understood. In the present study, we performed in vitro recording of abdominal ganglia cells of Aplysia juliana, and analyzed their burst firing patterns. We identified atypical bursting patterns dependent on temperature that were totally different from classical bursting patterns observed in R15 neurons of A. juliana. We classified these abnormal bursting patterns into type 1 and type 2; type 1 abnormal single bursts are composed of two kinds of spikes with a long interspike interval (ISI) followed by short ISI regular firing, while type 2 abnormal single bursts are composed of complex multiplets. To investigate the mechanism underlying the temperature dependence of abnormal bursting, we employed simulations using a modified Plant model and determined that the temperature dependence of type 2 abnormal bursting is related to temperature-dependent scaling factors and activation or inactivation of potassium or sodium channels.

Alteration of the haemocyte parameters, expression of immune‐related and neurohormone bursicon genes of giant river prawn Macrobrachium rosenbergii (de Man) in the response to salinity stress and ammonia stress

We examined the effects of salinity stress and ammonia stress on alteration of the haemocyte count, phenoloxidase activity, expressions of immune-related genes including prophenoloxidase (proPO), crustin, heat shock protein 70 (HSP70), and expressions of stress-responsive neurohormone (Bur-α and Bur-β) in the thoracic and abdominal ganglia of giant river prawn Macrobrachium. These parameters of prawn that subjected to salinity stress (transferred from 0‰ to 5‰ and 10‰), and subjected to ammonia-nitrogen (ammonia-N) stress (transferred from 0 to 0.262 mg/L and 0.786 mg/L) were examined after 0, 3, 6, 24, 72 and 168 hr respectively. During the initial period of 3, 6 and 24 hr, granulocyte haemocyte (granular and semi-granular hemocyte) count and PO activity significantly decreased, while expressions of Bur-α and Bur-β significantly increased. After 24 hr, granulocyte haemocyte count and PO activity significantly increased, whereas expressions of Bur-α and Bur-β significantly decreased. The expressions of proPO, crustin and HSP70 were significantly downregulated in the prawn that subjected to salinity stress and ammonia-N stress at all time periods of 3–168 hr. In conclusion, changes in the granulocyte haemocyte count of M. rosenbergii following salinity stress and ammonia-N stress are closely associated with the changes of Bur-α and Bur-β expressions.

Expression Profiling, Downstream Signaling, and Inter-subunit Interactions of GPA2/GPB5 in the Adult Mosquito Aedes aegypti.

GPA2/GPB5 and its receptor constitute a glycoprotein hormone-signaling system native to the genomes of most vertebrate and invertebrate organisms. Unlike the well-studied gonadotropins and thyrotropin, the exact function of GPA2/GPB5 remains elusive, and whether it elicits its functions as heterodimers, homodimers or as independent monomers remains unclear. Here, the glycoprotein hormone signaling system was investigated in adult mosquitoes, where GPA2 and GPB5 subunit expression was mapped and modes of its signaling were characterized. In adult Aedes aegypti mosquitoes, GPA2 and GPB5 transcripts co-localized to bilateral pairs of neuroendocrine cells, positioned within the first five abdominal ganglia of the central nervous system. Unlike GPA2/GPB5 homologs in human and fly, GPA2/GPB5 subunits in A. aegypti lacked evidence of heterodimerization. Rather, cross-linking analysis to determine subunit interactions revealed A. aegypti GPA2 and GPB5 subunits may form homodimers, although treatments with independent subunits did not demonstrate receptor activity. Since mosquito GPA2/GPB5 heterodimers were not evident by heterologous expression, a tethered fusion construct was generated for expression of the subunits as a single polypeptide chain to mimic heterodimer formation. Our findings revealed A. aegypti LGR1 elicited constitutive activity with elevated levels of cAMP. However, upon treatment with recombinant tethered GPA2/GPB5, an inhibitory G protein (Gi/o) signaling cascade is initiated and forskolin-induced cAMP production is inhibited. These results further support the notion that heterodimerization is a requirement for glycoprotein hormone receptor activation and provide novel insight to how signaling is achieved for GPA2/GPB5, an evolutionary ancient neurohormone.

Neural circuitry linking mating and egg laying in Drosophila females.

Mating and egg laying are tightly cooordinated events in the reproductive life ofall oviparous females. Oviposition is typically rare in virgin females but is initiated after copulation. Here we identify the neural circuitry that links egg laying to mating status in Drosophila melanogaster. Activation of female-specific oviposition descending neurons (oviDNs) is necessary and sufficient for egg laying, and is equally potent in virgin and mated females. After mating, sex peptide-a protein from the male seminal fluid-triggers many behavioural and physiological changes in the female, including the onset of egg laying1. Sex peptide is detected by sensory neurons in the uterus2-4, and silences these neurons and their postsynaptic ascending neurons in the abdominal ganglion5. We show that theseabdominal ganglion neurons directly activate the female-specific pC1 neurons. GABAergic (γ-aminobutyric-acid-releasing) oviposition inhibitory neurons (oviINs) mediate feed-forward inhibition from pC1 neurons to both oviDNs and their major excitatory input, the oviposition excitatory neurons (oviENs). By attenuating the abdominal ganglion inputs to pC1 neurons and oviINs, sex peptide disinhibits oviDNs to enable egg laying after mating. This circuitry thus coordinates the two key events in female reproduction: mating and egg laying.

Evolution and Comparative Physiology of Luqin-Type Neuropeptide Signaling.

Luqin is a neuropeptide that was discovered and named on account of its expression in left upper quadrant cells of the abdominal ganglion in the mollusc Aplysia californica. Subsequently, luqin-type peptides were identified as cardio-excitatory neuropeptides in other molluscs and a cognate receptor was discovered in the pond snail Lymnaea stagnalis. Phylogenetic analyses have revealed that orthologs of molluscan luqin-type neuropeptides occur in other phyla; these include neuropeptides in ecdysozoans (arthropods, nematodes) that have a C-terminal RYamide motif (RYamides) and neuropeptides in ambulacrarians (echinoderms, hemichordates) that have a C-terminal RWamide motif (RWamides). Furthermore, precursors of luqin-type neuropeptides typically have a conserved C-terminal motif containing two cysteine residues, although the functional significance of this is unknown. Consistent with the orthology of the neuropeptides and their precursors, phylogenetic and pharmacological studies have revealed that orthologous G-protein coupled receptors (GPCRs) mediate effects of luqin-type neuropeptides in spiralians, ecdysozoans, and ambulacrarians. Luqin-type signaling originated in a common ancestor of the Bilateria as a paralog of tachykinin-type signaling but, unlike tachykinin-type signaling, luqin-type signaling was lost in chordates. This may largely explain why luqin-type signaling has received less attention than many other neuropeptide signaling systems. However, insights into the physiological actions of luqin-type neuropeptides (RYamides) in ecdysozoans have been reported recently, with roles in regulation of feeding and diuresis revealed in insects and roles in regulation of feeding, egg laying, locomotion, and lifespan revealed in the nematode Caenorhabditis elegans. Furthermore, characterization of a luqin-type neuropeptide in the starfish Asterias rubens (phylum Echinodermata) has provided the first insights into the physiological roles of luqin-type signaling in a deuterostome. In conclusion, although luqin was discovered in Aplysia over 30 years ago, there is still much to be learnt about luqin-type neuropeptide signaling. This will be facilitated in the post-genomic era by the emerging opportunities for experimental studies on a variety of invertebrate taxa.

Engineering microscale systems for fully autonomous intracellular neural interfaces

Conventional electrodes and associated positioning systems for intracellular recording from single neurons in vitro and in vivo are large and bulky, which has largely limited their scalability. Further, acquiring successful intracellular recordings is very tedious, requiring a high degree of skill not readily achieved in a typical laboratory. We report here a robotic, MEMS-based intracellular recording system to overcome the above limitations associated with form factor, scalability, and highly skilled and tedious manual operations required for intracellular recordings. This system combines three distinct technologies: (1) novel microscale, glass–polysilicon penetrating electrode for intracellular recording; (2) electrothermal microactuators for precise microscale movement of each electrode; and (3) closed-loop control algorithm for autonomous positioning of electrode inside single neurons. Here we demonstrate the novel, fully integrated system of glass–polysilicon microelectrode, microscale actuators, and controller for autonomous intracellular recordings from single neurons in the abdominal ganglion of Aplysia californica (n = 5 cells). Consistent resting potentials (<−35 mV) and action potentials (>60 mV) were recorded after each successful penetration attempt with the controller and microactuated glass–polysilicon microelectrodes. The success rate of penetration and quality of intracellular recordings achieved using electrothermal microactuators were comparable to that of conventional positioning systems. Preliminary data from in vivo experiments in anesthetized rats show successful intracellular recordings. The MEMS-based system offers significant advantages: (1) reduction in overall size for potential use in behaving animals, (2) scalable approach to potentially realize multi-channel recordings, and (3) a viable method to fully automate measurement of intracellular recordings. This system will be evaluated in vivo in future rodent studies.

CAPA neuropeptides and their receptor form an anti-diuretic hormone signaling system in the human disease vector, Aedes aegypti

Insect CAPA neuropeptides are homologs of mammalian neuromedin U and are known to influence ion and water balance by regulating the activity of the Malpighian ‘renal’ tubules (MTs). Several diuretic hormones are known to increase primary fluid and ion secretion by insect MTs and, in adult female mosquitoes, a calcitonin-related peptide (DH31) called mosquito natriuretic peptide, increases sodium secretion to compensate for the excess salt load acquired during blood-feeding. An endogenous mosquito anti-diuretic hormone was recently described, having potent inhibitory activity against select diuretic hormones, including DH31. Herein, we functionally deorphanized, both in vitro and in vivo, a mosquito anti-diuretic hormone receptor (AedaeADHr) with expression analysis indicating highest enrichment in the MTs where it is localized within principal cells. Characterization using a heterologous in vitro system demonstrated the receptor was highly sensitive to mosquito CAPA neuropeptides while in vivo, AedaeADHr knockdown abolished CAPA-induced anti-diuretic control of DH31-stimulated MTs. CAPA neuropeptides are produced within a pair of neurosecretory cells in each of the abdominal ganglia, whose axonal projections innervate the abdominal neurohaemal organs, where these neurohormones are released into circulation. Lastly, pharmacological inhibition of nitric oxide synthase (NOS) and protein kinase G (PKG) signaling eliminated anti-diuretic activity of CAPA, highlighting the role of the second messenger cGMP and NOS/PKG in this anti-diuretic signaling pathway.

Biomechanical Stresses Due to Tissue Micromotion at the Neural Interface Modulate Intracellular Membrane Potentials

Micromotion in brain tissue at a neural interface can arise from respiration and vascular pulsation besides behavioral perturbations. We have reported 2-4 µm of brain tissue displacements in rodents due to vascular pulsation and 10-30 µm due to respiration. It has been hypothesized that the periodic membrane potential (MP) changes observed in neuronal intracellular recordings correlated to tissue micromotion were an artifact. In this study, using penetrating microelectrodes for intracellular recordings in an Aplysia californica abdominal ganglion, we show that the membrane potentials and firing rates change due to periodic cell compression. We show that these membrane potential changes are modulated by calcium, potassium, and mechanosensitive channel blockers. There were no membrane potential changes in response to periodic stresses <1.5 kPa in n=5 cells. Increases in amplitude of periodic stresses resulted in a linear increase in MP fluctuations; action potentials were generated in response to stresses >3 kPa for n=2 silent neurons suggesting the role of mechanosensitive receptors. Neurons that were already firing changed their firing rate in n=3 cells indicating that MP fluctuations are physiological and not artifacts. Application of 5-HT to the ganglion cell, resulted in decreased firing rate indicating that MP fluctuations are potentially modulated by mechanosensitive SK-channels in Aplysia. Preliminary in vivo rodent experiments show that periodic, intracellular membrane potentials fluctuate in the order of 10-30 mV. Using a custom double barrel pipette to (1) record neural activity and (2) to deliver drugs locally, these periodic membrane potential changes due to breathing and heart rate are inhibited by the mechanosensitive ion channel blocker gadolinium chloride. Results of this study suggest that cells at the electrode-brain tissue interface could be potentially undergoing chronic sub-threshold neuromodulation that could impact its long-term functionality and viability.

Duality of 5-HT Effects on Crayfish Motoneurons

Serotonin (5-HT) is a major neuromodulator acting on the nervous system. Its various effects have been studied in vertebrates, as well as in arthropods, from the cellular and subcellular compartments up to the behavioral level, which includes the control of mood, aggression, locomotion, and anxiety. The diversity of responses of neurons to 5-HT has been related to its mode of application, the diversity of 5-HT-receptors, and the animals’ social status history. In the locomotor network of socially isolated crayfish, the duality of 5-HT-evoked responses (excitatory/inhibitory) on motoneurons (MNs), sensorimotor pathways, and their consequences on motor network activity has largely been studied. The aim of the present report is to examine if this duality of exogenous 5-HT-evoked responses in the crayfish locomotor network can be reproduced by direct activation of 5-HT neurons in the case of socially isolated animals. Our previous studies have focused on the mechanisms supporting these opposite effects on MNs, pointing out spatial segregation of 5-HT receptors responsible either for positive or negative responses. Here, we report new findings indicating that excitatory and inhibitory effects can be achieved simultaneously in different leg MNs by the activation of a single 5-HT cell in the first abdominal ganglion.

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.

The time-dependent diffusivity in the abdominal ganglion of Aplysia californica: experiments and simulations

The nerve cells of the Aplysia are much larger than mammalian neurons. Using the Aplysia ganglia to study the relationship between the cellular structure and the diffusion MRI signal can potentially shed light on this relationship for more complex organisms. We measured the dMRI signal of chemically-fixed abdominal ganglia of the Aplysia at several diffusion times. At the diffusion times measured and observed at low b-values, the dMRI signal is mono-exponential and can be accurately represented by the parameter ADC (Apparent Diffusion Coefficient). We performed numerical simulations of water diffusion for the large cell neurons in the abdominal ganglia after creating geometrical configurations by segmenting high resolution T2-weighted (T2w) images to obtain the cell outline and then incorporating a manually generated nucleus. The results of the numerical simulations validate the claim that water diffusion in the large cell neurons is in the short diffusion time regime at our experimental diffusion times. Then, using the analytical short time approximation (STA) formula for the ADC, we showed that in order to explain the experimentally observed behavior, it is necessary to consider the nucleus and the cytoplasm as two separate diffusion compartments. By using a two compartment STA model, we were able to illustrate the effect of the highly irregular shape of the cell nucleus on the ADC.

Use of Transcriptomic Analysis for the Design of Antibodies to Serve as Neuronal and Synaptic Markers in Macrobrachium crustaceans

Crustaceans are major constituents of aquatic ecosystems and, as such, changes in their behavior and the structure and function of their bodies can serve as indicators of alterations in their immediate environment, such as those associated with climate change and anthropogenic contamination. We want to develop markers of crustacean neural circuits and terminal synaptic structure that can be used for quantifiable characterization with confocal microscopy. When compared with other invertebrate species such as Drosophila and C. elegans, little is known regarding crustacean genomic information, particularly of the animal's nervous system. We have used bioinformatics and a de novo transcriptome assembly approach to identify potential targets for developing specific antibodies to serve as nervous system markers for freshwater prawns of the Macrobrachium spp. These species of prawn are found in rivers of Puerto Rico and are being used to study the impact of exposure to emergent contaminants, such as phthalates and metabolites of pharmaceuticals, in natural sources of freshwater near urban areas.Total RNA was extracted from brain ganglia of Macrobrachium carcinus freshwater prawns and Illumina Next Generation Sequencing was performed using an Eel Pond mRNA Seq Protocol to construct a de novo transcriptome. Transcript sequences related to glutamate receptors were ran through multiple sequence conservation analysis using the MUSCLE webserver. Conserved sequences were analyzed for antigenicity to design specific antibodies. Raw data from the Illumina Sequencing yielded 97,202,662 sequences: 47,630,546 paired and 1,941,570 singletons. Assembly with Trinity resulted in 197,898 assembled sequences, of which 30,576 were annotated: 9,600 annotated by orthology, 17,197 annotated by homology, and 3,779 annotated by transcript families. In the search for glutamate receptor sequences, we found 138 sequences related to ionotropic receptors, 32 sequences related to metabotropic receptors, and 18 sequences to unidentified receptors. Using this information, we were able to develop antibodies for prawn AMPA ionotropic glutamate receptor 1 (GluRIA1), metabotropic glutamate receptor subunit 1 and 4 (mGluRG1S1 and mGluRG3S4), and NMDA ionotropic receptor 2B (GluRNMDA2B). Immunohistochemistry of the ventral nerve cord has thus far shown both punctuate and cell body staining of mGluRG1S1 located throughout all the central nervous system, including the eyestalks, brain, and the circumesophageal, subesophageal, thoracic and abdominal ganglia. The other antibodies are presently being tested in the prawn central nervous system as well as in peripheral synapses such as the neuromuscular junction.Support or Funding InformationNSF DBI‐1337284NSF CREST NSF HRD‐1736019NIH NIMHD 8G12‐MD007600 (RCMI)NIGMS‐RISE R25 GM061838 NSF (USA): CREST HRD‐1137725This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Distribution of neuropeptide F in the ventral nerve cord and its possible role on testicular development and germ cell proliferation in the giant freshwater prawn, Macrobrachium rosenbergii.

Neuropeptide F in invertebrates is a homolog of neuropeptide Y in mammals and it is a member of FMRFamide-related peptides. In arthropods, such as insects, there are two types of neuropeptide F comprising long neuropeptide F (NPF) and short neuropeptide F (sNPF). Both NPFs are known to play a crucial role in the regulations of foraging, feeding-related behaviors, circadian rhythm, stress responses, aggression and reproduction in invertebrates. We have earlier found that in the giant freshwater prawn, Macrobrachium rosenbergii, there are three isoforms of NPF and four isoforms of sNPF and that NPFs are expressed in the eyestalks and brain. In the present study, we investigate further the tissue distribution of NPF-I in the ventral nerve cord (VNC) and its role in the development of testes in small male (SM) Macrobrachium rosenbergii. By immunolocalization, using the rabbit polyclonal antibody against NPF-I as a probe, we could detect NPF-I immunoreactivity in the neuropils and neuronal clusters of the subesophageal ganglia (SEG), thoracic ganglia (TG) and abdominal ganglia (AG) of the SM prawns. In functional assays, the administrations of synthetic NPF-I (KPDPTQLAAMADALKYLQELDKYYSQVSRPRFamide) and sNPF (APALRLRFamide) peptides significantly increased the growth rates of SM prawns and significantly increased the gonadosomatic index (GSI) and proliferations of early germ cells in the seminiferous tubules of their testes. It is, therefore, suggestive that NPFs may play critical roles in energy homeostasis towards promoting growth as well as testicular development in prawns that could be applied in the aquaculture of this species.

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.

A population of descending tyraminergic/octopaminergic projection neurons of the insect deutocerebrum.

In this study, we describe a cluster of tyraminergic/octopaminergic neurons in the lateral dorsal deutocerebrum of desert locusts (Schistocerca gregaria) with descending axons to the abdominal ganglia. In the locust, these neurons synthesize octopamine from tyramine stress-dependently. Electrophysiological recordings in locusts reveal that they respond to mechanosensory touch stimuli delivered to various parts of the body including the antennae. A similar cluster of tyraminergic/octopaminergic neurons was also identified in the American cockroach (Periplaneta americana) and the pink winged stick insect (Sipyloidea sipylus). It is suggested that these neurons release octopamine in the ventral nerve cord ganglia and, most likely, convey information on arousal and/or stressful stimuli to neuronal circuits thus contributing to the many actions of octopamine in the central nervous system.

Distribution of abalone egg-laying hormone-like peptide in the central nervous system and reproductive tract of the male mud crab, Scylla olivacea

The mud crab, Scylla olivacea, is a high value economic marine animal in Thailand. However, collection of these crabs from natural habitat for local consumption and export has caused rapid population decline. Hence, aquaculture of this species is required and to this measure understanding of endocrine control of their reproduction must be understood. Egg laying hormone (ELH) is a neuropeptide synthesized by the bag cells (neurons) in the abdominal ganglia of Aplysia gastropods. It plays a critical role in controlling egg production and laying in gastropods, and its possible homolog (ELH-like peptide) was reported in the neural and ovarian tissues of prawns and recently in female reproductive tract of the blue swimming crab, Portunus pelagicus. In this study, we have studied the histology of the male reproductive tract in Scylla olivacea which are comprised of anterior testis, posterior testis, early proximal spermatic duct (ePSD), proximal spermatic duct (PSD), middle spermatic duct (MSD) and distal spermatic duct (DSD), by immunohistochemistry, detected an abalone ELH- immunoreactivity (aELH-ir) in epithelium of ducts in posterior testis and epithelium of all parts of spermatic duct. Furthermore, we could detect aELH-ir in neurons of cluster 9, 11, olfactory neuropil (ON) in the brain and in the small neurons located between the third and the fourth thoracic neuropils (T3-T4) and between the fourth and the fifth thoracic neuropils (T4-T5) of thoracic ganglia. Thus, the presence of aELH in male S. olivacea was designated the role of female egg laying behavior in the male mud crab.

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.

Modulation of Drosophila post-feeding physiology and behavior by the neuropeptide leucokinin.

Behavior and physiology are orchestrated by neuropeptides acting as central neuromodulators and circulating hormones. An outstanding question is how these neuropeptides function to coordinate complex and competing behaviors. In Drosophila, the neuropeptide leucokinin (LK) modulates diverse functions, but mechanisms underlying these complex interactions remain poorly understood. As a first step towards understanding these mechanisms, we delineated LK circuitry that governs various aspects of post-feeding physiology and behavior. We found that impaired LK signaling in Lk and Lk receptor (Lkr) mutants affects diverse but coordinated processes, including regulation of stress, water homeostasis, feeding, locomotor activity, and metabolic rate. Next, we sought to define the populations of LK neurons that contribute to the different aspects of this physiology. We find that the calcium activity in abdominal ganglia LK neurons (ABLKs), but not in the two sets of brain neurons, increases specifically following water consumption, suggesting that ABLKs regulate water homeostasis and its associated physiology. To identify targets of LK peptide, we mapped the distribution of Lkr expression, mined a brain single-cell transcriptome dataset for genes coexpressed with Lkr, and identified synaptic partners of LK neurons. Lkr expression in the brain insulin-producing cells (IPCs), gut, renal tubules and chemosensory cells, correlates well with regulatory roles detected in the Lk and Lkr mutants. Furthermore, these mutants and flies with targeted knockdown of Lkr in IPCs displayed altered expression of insulin-like peptides (DILPs) and transcripts in IPCs and increased starvation resistance. Thus, some effects of LK signaling appear to occur via DILP action. Collectively, our data suggest that the three sets of LK neurons have different targets, but modulate the establishment of post-prandial homeostasis by regulating distinct physiological processes and behaviors such as diuresis, metabolism, organismal activity and insulin signaling. These findings provide a platform for investigating feeding-related neuroendocrine regulation of vital behavior and physiology.

Intermediate-term memory in Aplysia involves neurotrophin signaling, transcription, and DNA methylation

Long-term but not short-term memory and synaptic plasticity in many brain areas require neurotrophin signaling, transcription, and epigenetic mechanisms including DNA methylation. However, it has been difficult to relate these cellular mechanisms directly to behavior because of the immense complexity of the mammalian brain. To address that problem, we and others have examined numerically simpler systems such as the hermaphroditic marine mollusk Aplysia californica. As a further simplification, we have used a semi-intact preparation of the Aplysia siphon withdrawal reflex in which it is possible to relate cellular plasticity directly to behavioral learning. We find that inhibitors of neurotrophin signaling, transcription, and DNA methylation block sensitization and classical conditioning beginning ∼1 h after the start of training, which is in the time range of an intermediate-term stage of plasticity that combines elements of short- and long-term plasticity and may form a bridge between them. Injection of decitabine (an inhibitor of DNA methylation that may have other actions in these experiments) into an LE sensory neuron blocks the neural correlates of conditioning in the same time range. In addition, we found that both DNA and RNA methylation in the abdominal ganglion are correlated with learning in the same preparations. These results begin to suggest the functions and integration of these different molecular mechanisms during behavioral learning.

Bursicon homodimers induce innate immune by activating the expression of anti-microbial peptide genes in the shrimp Neocaridina heteropoda

Bursicon is a neurohormone belonging to the cystine knot protein family. It consists of two subunits (burs α and burs β) and plays a pivotal role in cuticle tanning and wing expansion in insects. Recent studies show that homologous crustacean bursicon stimulates cuticle thickening and granulation of hemocytes in the crab Callinectes sapidus. Here we investigate whether bursicon homodimers function in immunoprotective defense systems of shrimp. We found that abdominal ganglion was the main neurohemal release site of bursicon in Neocaridina heteropoda. Bacterial infections induced overexpression of burs α (bursicon α) and burs β (bursicon β). RNAi of burs α, burs β or both inhibited the expression of anti-microbial peptide (AMP) genes. Treating shrimp adults with r-bursicon (recombinant bursicon) homodimers led to up-regulation of three AMP genes. Besides, through the induced AMPs, r-bursicon homodimers enhanced the bacteriostasis of shrimp in vivo and in vitro. These findings demonstrate a novel function of bursicon in crustacean that it induces innate immune via up-regulating the expression of genes encoding AMPs.