Suboesophageal Ganglion Research Articles

Melatonin in the seasonal response of the aphid Acyrthosiphon pisum.

Aphids display life cycles largely determined by the photoperiod. During the warm long-day seasons, most aphid species reproduce by viviparous parthenogenesis. The shortening of the photoperiod in autumn induces a switch to sexual reproduction. Males and sexual females mate to produce overwintering resistant eggs. In addition to this full life cycle (holocycle), there are anholocyclic lineages that do not respond to changes in photoperiod and reproduce continuously by parthenogenesis. The molecular or hormonal events that trigger the seasonal response (i.e., induction of the sexual phenotypes) are still unknown. Although circadian synthesis of melatonin is known to play a key role in vertebrate photoperiodism, the involvement of the circadian clock and/or of the hormone melatonin in insect seasonal responses is not so well established. Here we show that melatonin levels in the aphid Acyrthosiphon pisum are significantly higher in holocyclic aphids reared under short days than under long days, while no differences were found between anholocyclic aphids under the same conditions. We also found that melatonin is localized in the aphid suboesophageal ganglion (SOG) and in the thoracic ganglionic mass (TGM). In analogy to vertebrates, insect-type arylalkylamine N-acetyltransferases (i-AANATs) are thought to play a key role in melatonin synthesis. We measured the expression of four i-AANAT genes identified in A. pisum and localized two of them in situ in the insect central nervous systems (CNS). Levels of expression of these genes were compatible with the quantities of melatonin observed. Moreover, like melatonin, expression of these genes was found in the SOG and the TGM.

The neuroanatomy of the siboglinid Riftia pachyptila highlights sedentarian annelid nervous system evolution.

Tracing the evolution of the siboglinid group, peculiar group of marine gutless annelids, requires the detailed study of the fragmentarily explored central nervous system of vestimentiferans and other siboglinids. 3D reconstructions of the neuroanatomy of Riftia revealed that the “brain” of adult vestimentiferans is a fusion product of the supraesophageal and subesophageal ganglia. The supraesophageal ganglion-like area contains the following neural structures that are homologous to the annelid elements: the peripheral perikarya of the brain lobes, two main transverse commissures, mushroom-like structures, commissural cell cluster, and the circumesophageal connectives with two roots which give rise to the palp neurites. Three pairs of giant perikarya are located in the supraesophageal ganglion, giving rise to the paired giant axons. The circumesophageal connectives run to the VNC. The subesophageal ganglion-like area contains a tripartite ventral aggregation of perikarya (= the postoral ganglion of the VNC) interconnected by the subenteral commissure. The paired VNC is intraepidermal, not ganglionated over most of its length, associated with the ciliary field, and comprises the giant axons. The pairs of VNC and the giant axons fuse posteriorly. Within siboglinids, the vestimentiferans are distinguished by a large and considerably differentiated brain. This reflects the derived development of the tentacle crown. The tentacles of vestimentiferans are homologous to the annelid palps based on their innervation from the dorsal and ventral roots of the circumesophageal connectives. Neuroanatomy of the vestimentiferan brains is close to the brains of Cirratuliiformia and Spionida/Sabellida, which have several transverse commissures, specific position of the giant somata (if any), and palp nerve roots (if any). The palps and palp neurite roots originally developed in all main annelid clades (basally branching, errantian and sedentarian annelids), show the greatest diversity in their number in sedentarian species. Over the course of evolution of Sedentaria, the number of palps and their nerve roots either dramatically increased (as in vestimentiferan siboglinids) or were lost.

The functional connectivity between the locust leg pattern generators and the subesophageal ganglion higher motor center

Higher motor centers and central pattern generators (CPGs) interact in the control of coordinated leg movements during locomotion throughout the animal kingdom. The subesophageal ganglion (SEG) is one of the insect head ganglia reported to have a role in the control of walking behavior. Here we explored the functional relations between the SEG and the thoracic leg CPGs in the desert locust. Backfill staining revealed about 300 SEG descending interneurons (DINs) altogether. Recordings from an in-vitro isolated chain of thoracic ganglia, with intact or severed connections to the SEG, during pharmacological activation were used to determine how the SEG affects the centrally generated motor output to the legs. The SEG was demonstrated to both activate leg CPGs and synchronize their bilateral activity. The role of the SEG in insect locomotion is discussed in light of these findings.

Mass Spectrometric Quantification of Arousal Associated Neurochemical Changes in Single Honey Bee Brains and Brain Regions.

Honey bee foragers show a strong diurnal rhythm of foraging activity, and such behavioral changes are likely under the control of specific neuromodulators. To identify and quantify neuromodulators involved in regulating rest and arousal in honey bees, we established a mass spectrometric method for quantifying 14 different neurochemicals and precursor molecules. We measured forager type and brain region specific differences in amine levels from individual honey bee brains and brain regions. The observed differences in amine levels between resting and aroused foragers resemble findings in other species indicating a conserved molecular mechanism by glutamate and GABA in regulating arousal. Subesophageal ganglion specific changes in the histaminergic system and global increases in aspartate during arousal suggest a possible role of histamine and aspartate in feeding and arousal, respectively. More aminergic systems were significantly affected due to arousal in nectar foragers than in pollen foragers, implying that forager phenotypes differ not only in their food preference but also in their neuromodulatory signaling systems (brain states). Finally, we found that neurotransmitter precursors were better at distinguishing brain states in the central brain, while their end products correlated with arousal associated changes in sensory regions like the optic and antennal lobes.

Identification and expression of prostaglandin E synthase (PGES) gene in the central nervous system and ovary during ovarian maturation of the female mud crab, Scylla olivacea

Prostaglandins have important physiological roles in marine invertebrates, including larval development and reproduction. The prostaglandin E concentration fluctuates during the ovarian development of crustaceans. The biosynthetic pathway of prostaglandin, however, has not been well studied in portunid crabs, including in the mud crab, Scylla olivacea. In this study, the aim was to investigate the presence of prostaglandin E synthase (PGES), enzyme that catalyzes the terminal conversion in the prostaglandin E2 (PGE2) biosynthesis, and its gene expression in the central nervous system (CNS) and ovary during ovarian maturation of S. olivacea. cDNA sequence encoding PGES was cloned from the S. olivacea ovary. The PGES transcript of S. olivacea (Scyol-PGES) consists of 1258 nucleotides, which encodes for 420 amino acid PGES protein precursor. Investigation of gene expression by RT-PCR indicated that Scyol-PGES was detected in all organs studied. Based on in situ hybridization, Scyol-PGES was detected in the I to III stages for oocyte development of Stage 3 of ovarian development, and in the CNS, including the various neuronal clusters of the brain. In the ventral nerve cord, the Scyol-PGES gene was expressed in the neurons within the subesophageal, thoracic and abdominal ganglia. The Scyol-PGES gene expression as indicated by relative abundance of mRNA in the Stage 4 of ovarian development was greater than that at Stages 1 to 3 of ovarian development. This is the first report on PGES in the mud crab, S. olivacea, and its gene expression suggested the involvement of PGES in the ovarian development of this species.

A comparative analysis of the ventral nerve cord of Lithobius forficatus (Lithobiomorpha): morphology, neuroanatomy, and individually identifiable neurons

In light of competing hypotheses on arthropod phylogeny, independent data are needed in addition to traditional morphology and modern molecular approaches. One promising approach involves comparisons of structure and development of the nervous system. In addition to arthropod brain and ventral nerve cord morphology and anatomy, individually identifiable neurons (IINs) provide new character sets for comparative neurophylogenetic analyses. However, very few species and transmitter systems have been investigated, and still fewer species of centipedes have been included in such analyses. In a multi-methodological approach, we analyze the ventral nerve cord of the centipede Lithobius forficatus using classical histology, X-ray micro-computed tomography and immunohistochemical experiments, combined with confocal laser-scanning microscopy to characterize walking leg ganglia and identify IINs using various neurotransmitters. In addition to the subesophageal ganglion, the ventral nerve cord of L. forficatus is composed of the forcipular ganglion, 15 well-separated walking leg ganglia, each associated with eight pairs of nerves, and the fused terminal ganglion. Within the medially fused hemiganglia, distinct neuropilar condensations are located in the ventral-most domain. Immunoreactive neurons of different transmitter systems (allatostatin, histamine, and FMRF-amide) display serially homologous patterns that may lay the foundation for comparison with other arthropod taxa. Moreover, a pair of histaminergic neurons may constitute a promising intra- as well as interspecific IIN candidate.

Proprioceptive input to a descending pathway conveying antennal postural information: Terminal organisation of antennal hair field afferents

Like several other arthropod species, stick insects use their antennae for tactile exploration of the near-range environment and for spatial localisation of touched objects. More specifically, Carausius morosus continuously moves its antennae during locomotion and reliably responds to antennal contact events with directed movements of a front leg. Here we investigate the afferent projection patterns of antennal hair fields (aHF), proprioceptors known to encode antennal posture and movement, and to be involved in antennal movement control. We show that afferents of all seven aHF of C. morosus have terminal arborisations in the dorsal lobe (DL) of the cerebral (=supraoesophageal) ganglion, and descending collaterals that terminate in a characteristic part of the gnathal (=suboesophageal) ganglion. Despite differences of functional roles among aHF, terminal arborisation patterns show no topological arrangement according to segment specificity or direction of movement. In the DL, antennal motoneuron neurites show arborizations in proximity to aHF afferent terminals. Despite the morphological similarity of single mechanoreceptors of aHF and adjacent tactile hairs on the pedicel and flagellum, we find a clear separation of proprioceptive and exteroceptive mechanosensory neuropils in the cerebral ganglion. Moreover, we also find this functional separation in the gnathal ganglion.

Clinical Autogenous Transfers Into the Oocyte Donor’s Uterus of Embryos Produced by Intracytoplasmic sperm Injection

Prostaglandins have important physiological roles in marine invertebrates, including larval development and reproduction. The prostaglandin E concentration fluctuates during the ovarian development of crustaceans. The biosynthetic pathway of prostaglandin, however, has not been well studied in portunid crabs, including in the mud crab, Scylla olivacea. In this study, the aim was to investigate the presence of prostaglandin E synthase (PGES), enzyme that catalyzes the terminal conversion in the prostaglandin E2 (PGE2) biosynthesis, and its gene expression in the central nervous system (CNS) and ovary during ovarian maturation of S. olivacea. cDNA sequence encoding PGES was cloned from the S. olivacea ovary. The PGES transcript of S. olivacea (Scyol-PGES) consists of 1258 nucleotides, which encodes for 420 amino acid PGES protein precursor. Investigation of gene expression by RT-PCR indicated that Scyol-PGES was detected in all organs studied. Based on in situ hybridization, Scyol-PGES was detected in the I to III stages for oocyte development of Stage 3 of ovarian development, and in the CNS, including the various neuronal clusters of the brain. In the ventral nerve cord, the Scyol-PGES gene was expressed in the neurons within the subesophageal, thoracic and abdominal ganglia. The Scyol-PGES gene expression as indicated by relative abundance of mRNA in the Stage 4 of ovarian development was greater than that at Stages 1 to 3 of ovarian development. This is the first report on PGES in the mud crab, S. olivacea, and its gene expression suggested the involvement of PGES in the ovarian development of this species.

3D virtual histology at the host/parasite interface: visualisation of the master manipulator, Dicrocoelium dendriticum, in the brain of its ant host

Some parasites are able to manipulate the behaviour of their hosts to their own advantage. One of the most well-established textbook examples of host manipulation is that of the trematode Dicrocoelium dendriticum on ants, its second intermediate host. Infected ants harbour encysted metacercariae in the gaster and a non-encysted metacercaria in the suboesophageal ganglion (SOG); however, the mechanisms that D. dendriticum uses to manipulate the ant behaviour remain unknown, partly because of a lack of a proper and direct visualisation of the physical interface between the parasite and the ant brain tissue. Here we provide new insights into the potential mechanisms that this iconic manipulator uses to alter its host’s behaviour by characterising the interface between D. dendriticum and the ant tissues with the use of non-invasive micro-CT scanning. For the first time, we show that there is a physical contact between the parasite and the ant brain tissue at the anteriormost part of the SOG, including in a case of multiple brain infection where only the parasite lodged in the most anterior part of the SOG was in contact with the ant brain tissue. We demonstrate the potential of micro-CT to further understand other parasite/host systems in parasitological research.

The use of MEMRI for monitoring central nervous system activity during intact insect walking

BackgroundMonitoring neuronal activity in the intact behaving animal is most desired in neuroethological research, yet it is rarely straightforward or even feasible. Here we present the use of manganese enhanced magnetic resonance imaging (MEMRI), a technique allowing monitoring the activity of an animal’s nervous system during specific behavioral patterns. Using MEMRI we were able to show activity in different ganglia of the central nervous system of intact locusts during walking. ResultsWe injected two groups of locusts with manganese, which serves as a magnetic contrast agent. One group was forced to walk on a treadmill for two hours, while the other was immobilized and served as a control. Subsequently, all animals were scanned in a T1 MRI protocol, and the accumulation of manganese in the neuronal tissues that were active during walking was demonstrated by comparing the scans of the two groups. Two neuronal sites showed significantly higher T1 signal in the walking locusts compared to the immobilized ones: the prothoracic ganglion, which locally controls the front legs, and the subesophageal ganglion, a head ganglion which takes part in initiation and maintenance of walking. ConclusionMEMRI is a potent, non-invasive technique for monitoring neuronal activity in intact locusts, and arthropods in general. Specifically, it provides a promising way for revealing the role of central and high-order neuronal structures in motor behaviors such as walking.

Isolation and functional characterization of the pheromone biosynthesis activating neuropeptide receptor of Chinese oak silkworm, Antheraea pernyi

Insect pheromone biosynthesis activating neuropeptide (PBAN) controls the synthesis and actuating of sex pheromones of female adult. In the current examination, the full-length cDNA encoding the PBAN receptor was cloned from the pheromone gland (PG) of Antheraea pernyi (AntpePBANR). The AntpePBANR displayed the characteristic seven transmembrane areas of the G protein-coupled receptor (GPCR) and was closely related to the PBANR from Bombyx mori and Manduca sexta in the phylogenetic tree. The AntpePBANR expressed in mammalian cell lines were enacted by AntpePBAN in a concentration-dependent manner. AntpePBANR activation resulted in the calcium mobilization but did not activate the cAMP elevation pathway. Cells expressing AntpePBANR were profoundly responsive to Antpe-γ-SGNP (suboesophageal ganglion neuropeptides) and Antpe-DH (diapause hormone), different individuals from FXPRLamide (X = T, S or V) family in A. pernyi. Deletion of residues in the C-terminal hexapeptide (FSPRLamide) proved that P, R and L played the key parts in initiating the AntpePBANR, the amination to the last C terminal residues which can also likewise impact the activation of AntpePBAN receptor altogether. The mRNA of the AntpePBANR gene demonstrated the most noteworthy transcript levels in pheromone gland followed by fat body.

Motor innervation pattern of labral muscles of Locusta migratoria.

The current study investigates the motor innervation pattern of labral muscles in the adult locust and tries to interpret the results in the light of the hypothesis that the labrum phylogenetically developed by the fusion of paired appendages associated with the intercalary segment. Using Neurobiotin™ as a retrograde neuronal tracer, specific motor nerves or individual labral muscles were stained. Results show that the labral muscles receive innervation from tritocerebrum and suboesophageal ganglion. The axons of many motor neurons use three different pathways to cross the midline in the periphery to innervate ipsi- and contralateral muscles. Intracellular recordings from fibers of individual muscles and simultaneous recordings from motor neurons imply that the labral muscles lack inhibitory innervation. The location of motor neurons in both tritocerebrum and suboesophageal ganglion supports the notion that the labrum is innervated by the so-called intercalary segment. That many of the efferent axons cross the midline in the periphery might be explained by the hypothesis that the labrum derives from a fusion of appendages.

Comparison of ventral organ development across Pycnogonida (Arthropoda, Chelicerata) provides evidence for a plesiomorphic mode of late neurogenesis in sea spiders and myriapods

BackgroundComparative studies of neuroanatomy and neurodevelopment provide valuable information for phylogenetic inference. Beyond that, they reveal transformations of neuroanatomical structures during animal evolution and modifications in the developmental processes that have shaped these structures. In the extremely diverse Arthropoda, such comparative studies contribute with ever-increasing structural resolution and taxon coverage to our understanding of nervous system evolution. However, at the neurodevelopmental level, in-depth data remain still largely confined to comparably few laboratory model organisms. Therefore, we studied postembryonic neurogenesis in six species of the bizarre Pycnogonida (sea spiders), which – as the likely sister group of all remaining chelicerates – promise to illuminate neurodevelopmental changes in the chelicerate lineage.ResultsWe performed in vivo cell proliferation experiments with the thymidine analogs 5-bromo-2′-deoxyuridine and 5-ethynl-2′-deoxyuridine coupled to fluorescent histochemical staining and immunolabeling, in order to compare ventral nerve cord anatomy and to localize and characterize centers of postembryonic neurogenesis. We report interspecific differences in the architecture of the subesophageal ganglion (SEG) and show the presence of segmental “ventral organs” (VOs) that act as centers of neural cell production during gangliogenesis. These VOs are either incorporated into the ganglionic soma cortex or found on the external ganglion surface. Despite this difference, several shared features support homology of the two VO types, including (1) a specific arrangement of the cells around a small central cavity, (2) the presence of asymmetrically dividing neural stem cell-like precursors, (3) the migration of newborn cells along corresponding pathways into the cortex, and (4) the same VO origin and formation earlier in development.ConclusionsEvaluation of our findings relative to current hypotheses on pycnogonid phylogeny resolves a bipartite SEG and internal VOs as plesiomorphic conditions in pycnogonids. Although chelicerate taxa other than Pycnogonida lack comparable VOs, they are a characteristic feature of myriapod gangliogenesis. Accordingly, we propose internal VOs with neurogenic function to be part of the ground pattern of Arthropoda. Further, our findings illustrate the importance of dense sampling in old arthropod lineages – even if as gross-anatomically uniform as Pycnogonida – in order to reliably differentiate plesiomorphic from apomorphic neurodevelopmental characteristics prior to outgroup comparison.

The subesophageal ganglion modulates locust inter-leg sensory-motor interactions via contralateral pathways

The neural control of insect locomotion is distributed among various body segments. Local pattern-generating circuits at the thoracic ganglia interact with incoming sensory signals and central descending commands from the head ganglia. The evidence from different insect preparations suggests that the subesophageal ganglion (SEG) may play an important role in locomotion-related tasks. In a previous study, we demonstrated that the locust SEG modulates the coupling pattern between segmental leg CPGs in the absence of sensory feedback. Here, we investigated its role in processing and transmitting sensory information to the leg motor centers and mapped the major related neural pathways. Specifically, the intra- and inter-segmental transfer of leg-feedback were studied by simultaneously monitoring motor responses and descending signals from the SEG. Our findings reveal a crucial role of the SEG in the transfer of intersegmental, but not intrasegmental, signals. Additional lesion experiments, in which the intersegmental connectives were cut at different locations, together with double nerve staining, indicated that sensory signals are mainly transferred to the SEG via the connective contralateral to the stimulated leg. We therefore suggest that, similar to data reported for vertebrates, insect leg sensory-motor loops comprise contralateral ascending pathways to the head and ipsilateral descending ones.

A new black fly (Diptera: Simuliidae) species of the subgenusAsiosimulium Takaoka Choochote from Thailand.

A new black fly species of the subgenus Asiosimulium Takaoka Choochote of the genus Simulium was recognized from northeastern Thailand based on morphology, mitochondrial DNA and ecology. This black fly species has similar morphological characteristics to Simulium oblongum Takaoka and Choochote that was also described from the same geographic region in all life stages. However, this new species could be distinguished at the adult stage by coloration of the maxillary palp and in the larval stage by the presence of a pigmented subesophageal ganglion that is lacking in S. oblongum. Genetic distance and phylogenetic analyses based on mitochondrial cytochrome c oxidase I sequences clearly differentiated the two species with minimum genetic distance of 3.51%. These species are also ecologically isolated as S. oblongum is found only at low elevation (<650 m above sea level) but the new species occurs only at high elevation (>1,100 m above sea level).

Tyraminergic modulation of agonistic outcomes in crayfish.

Octopamine, a biogenic amine, modulates various behaviors, ranging from locomotion and aggression to learning and memory in invertebrates. Several studies recently demonstrated that tyramine, the biological precursor of octopamine, also affects behaviors independent of octopamine. Here we investigated the involvement of tyramine in agonistic interaction of the male crayfish Procambarus clarkii. When male crayfish fight, larger animals (3-7% difference in body length) are more likely to win. By contrast, direct injection of tyramine or octopamine counteracted the physical advantage of larger animals. Tyramine or octopamine-injected naive large animals were mostly beaten by untreated smaller naive animals. This pharmacological effect was similar to the loser effect in which subordinate larger animals are frequently beaten by smaller animals. Furthermore, loser effects were partly eliminated by either injection of epinastine, an octopamine blocker, or yohimbine, a tyramine blocker, and significantly diminished by injection of a mixture of both blockers. We also observed that tyramine levels in the subesophageal ganglion were remarkably increased in subordinate crayfish after losing a fight. These results suggest that tyramine modulates aggressive levels of crayfish and contributes to the loser effect in parallel with octopamine.

Independent suboesophageal neuronal innervation of the defense gland and longitudinal muscles in the stick insect (Peruphasma schultei) prothorax

This study investigates the neuroanatomy of the defense gland and a related muscle in the stick insect Peruphasma schultei with axonal tracing and histological sections. The gland is innervated by three neurons through the Nervus anterior of the suboesophageal ganglion (SOG), the ipsilateral neuron (ILN), the contralateral neuron (CLN) and the prothoracic intersegmental neuron (PIN). The ILN has a large soma which is typical for motoneurons that cause fast contraction of large muscles and its dendrites are located in motor-sensory and sensory neuropile areas of the SOG. The CLN might be involved in the coordination of bilateral or unilateral discharge as its neurites are closely associated to the ILN of the contralateral gland. Close to the ejaculatory duct of the gland lies a dorsal longitudinal neck muscle, musculus pronoto-occipitalis (Idlm2), which is likely indirectly involved in gland discharge by controlling neck movements and, therefore, the direction of discharge. This muscle is innervated by three ventral median neurons (VMN). Thus, three neuron types (ILN, CLN, and PIN) innervate the gland muscle directly, and the VMNs could aid secretion indirectly. The cytoanatomy of motorneurons innervating the defense gland and neck muscle are discussed regarding the structure and functions of the neuropile in the SOG. As a basis for the neuroanatomical study on the defense gland we assembled a map of the SOG in Phasmatodea.

Ampulexins: A New Family of Peptides in Venom of the Emerald Jewel Wasp, Ampulex compressa

The parasitoid wasp Ampulex compressa injects venom directly into the brain and subesophageal ganglion of the cockroach Periplaneta americana, inducing a 7 to 10 day lethargy termed hypokinesia. Hypokinesia presents as a significant reduction in both escape response and spontaneous walking. We examined aminergic and peptidergic components of milked venom with HPLC and MALDI-TOF mass spectrometry. HPLC coupled with electrochemical detection confirmed the presence of dopamine in milked venom, while mass spectrometry revealed that the venom gland and venom sac have distinct peptide profiles, with milked venom predominantly composed of venom sac peptides. We isolated and characterized novel α-helical, amphipathic venom sac peptides that constitute a new family of venom toxins termed ampulexins. Injection of the most abundant venom peptide, ampulexin 1, into the subesophageal ganglion of cockroaches resulted in a short-term increase in escape threshold. Neither milked venom nor venom peptides interfered with growth of Escherichia coli or Bacillus thuringiensis on agar plates, and exposure to ampulexins or milked venom did not induce cell death in Chinese hamster ovary cells (CHO-K1) or Hi5 cells ( Trichoplusia ni).

Multiple Identified Neurons and Peripheral Nerves Innervating the Prothoracic Defense Glands in Stick Insects Reveal Evolutionary Conserved and Novel Elements of a Chemical Defense System

The defense glands in the dorsal prothorax are an important autapomorphic trait of stick insects (Phasmatodea). Here, we study the functional anatomy and neuronal innervation of the defense glands in Anisomorpha paromalus (Westwood, 1859) (Pseudophasmatinae), a species which sprays its defense secretions when disturbed or attacked. We use a neuroanatomical approach to identify the nerves innervating the gland muscles and the motoneurons with axons in the different nerves. The defense gland is innervated by nerves originating from two segments, the suboesophageal ganglion and the prothoracic ganglion. Axonal tracing confirms the gland innervation via the anterior suboesophageal nerve, and two intersegmental nerves, the posterior suboesophageal nerve and the anterior prothoracic nerve. Axonal tracing of individual nerves reveals eight identified neuron types in the suboesophageal or prothoracic ganglion. The strongest innervating nerve of the gland is the anterior suboesophageal nerve, which also supplies dorsal longitudinal thorax muscles (neck muscles) by separate nerve branches. Tracing of individual nerve branches reveals different sets of motoneurons innervating the defense gland (one ipsilateral and one contralateral suboesophageal neuron) or the neck muscle (ventral median neurons). The ipsilateral and contralateral suboesophageal neurons have no homologs in related taxa like locusts and crickets, and thus evolved within stick insects with the differentiation of the defense glands. The overall innervation pattern suggests that the longitudinal gland muscles derived from dorsal longitudinal neck muscles. In sum, the innervating nerves for dorsal longitudinal muscles are conserved in stick insects, while the neuronal control system was specialized with conserved motoneurons for the persisting neck muscles, and evolutionarily novel suboesophageal and prothoracic motoneurons innervating the defense gland.

Identification and pheromonotropic activity of pheromone biosynthesis activating neuropeptide in Maruca vitrata

Pheromone biosynthesis activating neuropeptide (PBAN) produced in the suboesophageal ganglion stimulates pheromone biosynthesis in the pheromone gland, mediating sexual behaviors. The nucleotide sequences of PBAN and PBAN receptor (PBANr) in the legume pod borer, Maruca vitrata, were obtained from the transcriptome of the head and the pheromone gland, respectively. PBAN was expressed in all developmental stages. In contrast, PBANr was only detected in all developmental stages except for male adult. To examine the pheromonotropic activity of PBAN (Mvi-PBAN) in M. vitrata, Mvi-PBAN was chemically synthesized and injected into female adult. The pheromone production was increased in the dose-dependent manner and exhibited the maximal increase 2h post-injection.