The scolopidial accessory organ in the Jerusalem cricket (Orthoptera: Stenopelmatidae).

Multiple mechanosensory organs form the subgenual organ complex in orthopteroid insects, located in the proximal tibia. In several Ensifera (Orthoptera), a small chordotonal organ, the so-called accessory organ, is the most posterior part of this sensory complex. In order to document the presence of this accessory organ among the Ensifera, the chordotonal sensilla and their innervation in the posterior tibia of two species of Jerusalem crickets (Stenopelmatidae: Stenopelmatus) is described. The sensory structures were stained by axonal tracing. Scolopidial sensilla occur in the posterior subgenual organ and the accessory organ in all leg pairs. The accessory organ contains 10-17 scolopidial sensilla. Both groups of sensilla are commonly spatially separated. However, in few cases neuronal fibres occurred between both organs. The two sensillum groups are considered as separate organs by the general spatial separation and innervation by different nerve branches. A functional role for mechanoreception is considered: since the accessory organ is located closely under the cuticle, sensilla may be suited to detect vibrations transferred over the leg's surface. This study extends the known taxa with an accessory organ, which occurs in several taxa of Ensifera. Comparative neuroanatomy thus suggests that the accessory organ may be conserved at least in Tettigoniidea.

Histological and ultrastructural features of the rectum in Poecilimon cervus Karabag, 1950 (Orthoptera: Tettigoniidae).

The morphology and ultrastructure of the rectum in Poecilimon cervus Karabag, 1950 (Orthoptera, Tettigoniidae) were analyzed by light microscope, scanning (SEM) and transmission electron microscopes (TEM). The rectum is the final part of the digestive tract that plays an important role in water reabsorption in insects and so provides osmoregulation. In the transverse sections, six rectal pads and columnar epithelium can be distinguished. The cuticular intima lines the lumen at the apical side of the epithelium. In the cytoplasm, there are numerous mitochondria, some endocytic vesicles, secreting vesicles whose sizes differ according to the area in the cell, and a nucleus with globular in shape. With this study, we aimed to demonstrate the ultrastructure of the rectum of P. cervus and differences or similarities of with other species.

How many mechanosensory organs in the bushcricket leg? Neuroanatomy of the scolopidial accessory organ in Tettigoniidae (Insecta: Orthoptera).

The subgenual organ and associated scolopidial organs are well studied in Orthoptera and related taxa. In some insects, a small accessory organ or Nebenorgan is described posterior to the subgenual organ. In Tettigoniidae (Ensifera), the accessory organ has only been noted in one species though tibial sensory organs are well studied for neuroanatomy and physiology. Here, we use axonal tracing to analyse the posterior subgenual organ innervated by the main motor nerve. Investigating seven species from different groups of Tettigoniidae, we describe a small group of scolopidial sensilla (5-9 sensory neurons) which has features characteristic of the accessory organ: posterior tibial position, innervation by the main leg nerve rather than by the tympanal nerve, orientation of dendrites in proximal or ventro-proximal direction in the leg, and commonly association with a single campaniform sensillum. The neuroanatomy is highly similar between leg pairs. We show differences in the innervation in two species of the genus Poecilimon as compared to the other species. In Poecilimon, the sensilla of the accessory organ are innervated by one nerve branch together with the subgenual organ. The results suggest that the accessory organ is part of the sensory bauplan in the leg of Tettigoniidae and probably Ensifera.

Light and electron microscopy study of the spermatheca of Eupholidoptera chabrieri bimucronata (Ramme, 1927) and Uromenus brevicollis trinacriae La Greca 1964 (Orthoptera: Tettigoniidae).

A study by both optical and electron microscopy has been carried out on the spermatheca of Eupholidoptera chabrieri bimucronata and Uromenus brevicollis trinacriae (Orthoptera: Tettigoniidae). In both the examined species, the spermatheca consists of a sac/kidney-shaped seminal receptacle and a more or less tortuous spermathecal duct that opens into the common oviduct. The wall of both the organs consists of a pseudostratified epithelium surmounted by a cuticular intima; the latter is made up of a thicker endocuticle and an epicuticle. The epithelium shows two different cell types, irregularly arranged and with well differentiated functions: cuticle-forming and gland cells. In both the species, the cuticle-forming cells perform other functions, in addition to producing the cuticular intima. The gland cells never come in contact with the cuticular intima, have inside the reservoir a secretion whose appearance can diversify also in contiguous zones of the seminal receptacle. Based on our findings in both the species, the functions of the seminal receptacle would differ from those of the spermathecal duct. In the latter, some areas of the wall of the connecting tract show an activity of lysis, by contiguous epithelial cells, that could play a role in control and selection of spermatozoa. As for the feather-shaped spermatodesms, similar in both the species, freeze-fracture observations have shown that the acrosome of each spermatozoon regularly covers three-quarters of the extension of the acrosome of the following spermatozoon. Finally, the significance of our findings, compared with what is known in literature, is discussed.

The complete mitochondrial genome of Filchnerella beicki Ramme, 1931 (Orthoptera: Acridoidea: Pamphagidae).

Abstract The complete mitochondrial genome of Filchnerella beicki Ramme, 1931, which was collected from the Ningxia Hui Autonomous Region of China, is reported here. It is 15,658 bp in length and contains 72.3% AT. All Filchnerella beicki protein-coding sequences start with a typical ATN codon, excluding cox1. The usual termination codon (TAN) and incomplete stop codons (T) were found from 13 protein-coding genes. All tRNA genes could be folded into the typical cloverleaf secondary structure, excluding trnS(AGN) which forms another structure. The sizes of the large and small ribosomal RNA genes are 1315 and 853 bp, respectively. The AT content of the A+T-rich region is 83.1%.

Establishment and characterization of an embryonic cell line from Gampsocleis gratiosa (Orthoptera: Tettigoniidae).

The first continuous cell line from the embryo of Gampsocleis gratiosa (Orthoptera: Tettigoniidae), designated as RIRI-GG1, was established. This cell line was serially subcultured in modified Grace medium. The cells were grown adherent to a culture flask and had spindle-like and polygonal shapes. The chromosome number ranged from 26 to 79 at the 50th passage, and 68% of cells had a diploid chromosome number. The growth rate was determined at the 53rd passage, and the population doubling time was calculated to be 122.1 h. The rDNA internal transcribed spacer and the mitochondrial cytochrome c oxidase subunit I gene sequence analysis indicated that the RIRI-GG1 cell line was derived from G. gratiosa. This cell line had no apparent susceptibility to Autographa californica nucleopolyhedrovirus and Bombyx mori nucleopolyhedrovirus.

Role of the fat body in the cave crickets Troglophilus cavicola and Troglophilus neglectus (Rhaphidophoridae, Saltatoria) during overwintering.

The cave crickets Troglophilus cavicola and Troglophilus neglectus are the most widely distributed European species of the family Rhaphidophoridae. Their life cycles span two years. They overwinter twice in caves in 4-6 months lasting diapause, T. cavicola in warmer microhabitats. In caves, older T. cavicola undergo sexual maturation, while T. neglectus do not. We hypothesized that the use of energy-supplying compounds and reserve proteins in the fat body is more extensive in T. cavicola than in T. neglectus. We analyzed the contents and morphology of lipid droplets, glycogen rosettes and protein granula at the beginning, the middle and the end of overwintering applying optic, TEM and biochemical methods. In all individuals, the fat body is composed of about 40 oval ribbons consisted of gradually changing adipocytes and urocytes. T. cavicola use glycogen continuously, and stop using lipids in the middle of overwintering, while this is inverse in T. neglectus. Till the middle of overwintering, all individuals exploit proteins, afterwards they are unevenly exploited. We found that the fat body is differently engaged in metabolism of both cave crickets during overwintering, supporting a more glycogen-dependent metabolism in T. cavicola, and a more lipid-dependent one in T. neglectus.

Formation and rearrangement of spermatodesms in males of some Orthoptera Tettigoniidae.

In the male genital tract of Tettigoniidae, the spermatodesms are composed of a limited number of spermatozoa whose nuclei and acrosomes are covered by a mucous cap. The formation of the cap begins in the testicular cyst during the lengthening of the apical prolongations of the spermatids and the spermatids' simultaneous division into small bundles or spermatodesms. The cap material is formed from a loosely arranged material in the lumen of the cyst, probably produced by the secretory activity of the delimiting cells. Another characteristic aspect of the Tettigoniidae is the rearrangement of the cap inside the spermiduct that seems to start when material from the lumen of the organ enters from the basal part of the cap. Except for the fibrils of the peripheral lamina, the fibrils of the cap undergo a marked degradation process. The final organization of the spermatodesm cap is reached inside the seminal vesicles; it consists of the fibrillar peripheral lamina delimiting an interior made up of loosely arranged fibrils immersed in a finely granular matrix.

Phylogenetic reconstruction of the family Acrypteridae (Orthoptera: Acridoidea) based on mitochondrial cytochrome B gene.

Sequences from the mitochondrial cytochrome b gene (Cyt b) were determined for 25 species from the superfamily Acridoidae and the homologous sequences of 19 species of grasshoppers were downloaded from the GenBank data library. The purpose was to develop a molecular phylogeny of the Acrypteridae, and to interpret the phylogenetic position of the family within the superfamily Acridoidea. Phylogeny was reconstructed by Maximum-parsimony (MP) and Bayesian criteria using Yunnanites coriacea and Tagasta marginella as outgroups. The alignment length of the fragments was 384 bp after excluding ambiguous sites, including 167 parsimony informative sites. In the fragments, the percentages of A + T and G + C were 70.7% and 29.3%, respectively. The monophyly of Arcypteridae is not supported by phylogenetic trees. Within the Arcypteridae, neither Arcypterinae nor Ceracrinae is supported as a monophyletic group. The current genus Chorthippus is not a monophyletic group, and should be a polyphyletic group. The present results are significantly different from the classification scheme of Arcypteridae, which is based on morphology.

Embryonic development of Galloisiana yuasai Asahina, with special reference to external morphology (insecta: Grylloblattodea).

The embryogenesis of Grylloblattodea, one of the most primitive of the polyneopteran orders, is described using Galloisiana yuasai with special reference to external morphology. The egg membranes are characterized by an endochorion crossed by numerous vertical aeropyles and a fairly thin vitelline membrane, features shared by Mantophasmatodea. The inner layer formation is of the fault type. Serosal elements in the amnioserosal fold differentiate into hydropylar cells, to function in water absorption together with specialized amniotic structures, i.e., an amniotic strand and a thickened amnion. The germ band is of the short germ type. The germ band immerses deep into the yolk after its full elongation along the egg surface, and in this respect blastokinesis closely resembles that of Mantophasmatodea. The embryological features, i.e., those on egg membranes and blastokinesis, may suggest a closer affinity of Grylloblattodea and Mantophasmatodea. Appendages, ectodermal invaginations, and sternal and pleural sclerites are discussed in the light of serial homology, to provide a new basis for elucidating the insect body plan. Appendages are divided into the proximal coxopodite and distal telopodite, the former being divided further into the subcoxa and coxa. Subcoxal and coxal elements are identified in the mandible as well as in the abdominal appendages. The subcoxa is divided into the epimeron and episternum by the pleural suture in thoracic segments. Likewise, in the abdominal segments the subcoxa is divided into two, although the homologs of the epimeron and episternum are not sclerotized, and in the labial segment the subcoxal derivative or the postmentum is divided into the submentum and mentum. Two coxal endites bulge out from the medial side of the gnathal appendages. The mandibular molar and incisor, maxillary lacinia and galea, and labial glossa and paraglossa are serially homologous with each other. In the thoracic segments the original embryonic sternum or "protosternum" is largely replaced by subcoxal elements, and merely remains as a small anterior presternum and a posterior spinasternum. A major part of the venter is represented by the derivatives of the episternum such as an extensive basisternum, katepisternum, and trochantin and the medial element of the epimeron. The pleuron is derived from the episternal elements or the anepisternum and preepisternum, which bears a spiracle in the mesothorax and metathorax, and the lateral element of the epimeron. The homolog of the preepisternum in the prothorax is the cervical sclerite, but with no spiracle developed. A median ventral invagination arises in the thoracic segments as a spina, and the homolog of the spina develops into the eversible sac in the first abdominal segment.

The presence and distribution of crustacean cardioactive peptide in the central and peripheral nervous system of the stick insect, Baculum extradentatum.

Crustacean cardioactive peptide (CCAP)-like immunoreactivity was localized and quantified in the central and peripheral nervous system of the Vietnamese stick insect, Baculum extradentatum, using immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). The brain, frontal ganglion, suboesophageal ganglion and ventral nerve cord displayed neurons and processes with CCAP-like immunoreactivity. The brain, in comparison to the other parts of the central nervous system, contained the greatest amount of CCAP (167 +/- 18 fmol), and showed CCAP-like staining in neurons, neuropil regions and the central complex. There were also CCAP-like varicosities and processes associated with the corpus cardiacum. The alimentary canal of B. extradentatum contained CCAP with the largest amount localized in the midgut (1110 +/- 274 fmol CCAP equivalents). The midgut contained numerous endocrine-like cells which stained positively for CCAP, whereas the foregut and hindgut revealed an extensive network of CCAP-like immunoreactive axons and varicosities. Based on physiological assays, the hindgut of the stick insect was found to be sensitive to CCAP, showing dose-dependent increases in contractions with threshold at 10(-10) M CCAP and maximal response at 5 x 10(-7) M CCAP. There were negligible quantities of CCAP in the oviducts and no CCAP-like immunoreactivity was associated with the oviducts. CCAP had no effect on spontaneous contractions of the oviducts. The presence of CCAP in the central nervous system, the stomatogastric nervous system, the corpus cardiacum and the alimentary canal, suggest broad ranging roles for CCAP in B. extradentatum.

Somatotopic mapping of chordotonal organ neurons in a primitive ensiferan, the New Zealand tree weta Hemideina femorata: I. femoral chordotonal organ.

The femoral chordotonal organ (FCO) in orthopteran insects comprises several hundred sensory neurons, making it one of the most complex insect proprioceptors. The sensory neurons are suspended from the proximal femur, connecting distally to ligaments and to a needle-like apodeme extending from the proximal tibia. They monitor the position and movement of the tibia. To address how this complexity depends on evolutionary status and function, the morphology of the FCO neurons in the primitive orthopteran Hemideina femorata was investigated by staining small populations of identified afferents. As in crickets, the FCOs in all legs of the weta comprise partly fused ventral and dorsal scoloparia, with the former containing two groups of somata, the ventral group (VG) and the dorsal group (DG). However, the dendrites of the DG insert into thin connective tissue attached to the ventral side of the dorsal ligament, forming a "third scoloparium." The VG afferents terminate mainly in the motor association neuropils, whereas afferents from the dorsal scoloparium neurons terminate exclusively in the vibratory neuropil as do the afferents from the subgenual organ, a substrate vibration detector. Several afferents originating in the DG have extensive terminations in the motor association-, vibratory-, and auditory-processing neuropils, indicating lesser functional specialization than in the other groups. The evolutionary development of the FCO is discussed from a comparative viewpoint.

[Electron microscope study of haemolymph cells of Metrioptera roeselii (Orthoptera, Tettigoniidae)]. / Elektronno-mikroskopicheskoe issledovanie kletok gemolimfy skachka zelenogo Metrioptera roeselii (Orthoptera, Tettigoniidae).

On the basis of patterns of haemocyte ultrastructure and functions at preimago and imago stages of Metrioptera roeselii, secretory cells of granulocyte type were recognized in the haemolymph. The development of granulocytes was traced starting from their formation up to cell death and destruction. The haemocytes develop as "dark" and "light" cells, differing in their functional activity, although their ultrastructure is similar. In the cytoplasm of granulocytes, granules of both mitochondrial and nuclear origin were detected. Simultaneously two processes occur in the cells--the accumulation and discharge of granules.

Synaptonemal complex analysis of the X1X2Y trivalent in Mantis religiosa L. males: inferences on the origin and maintenance of the sex-determining mechanism.

Characterization of sex chromosomes in males of Mantis religiosa L. (2n = 24 + X1X2Y) was carried out by C-banding, silver staining and fluorescence in situ hybridization. They are meta- or submetacentric, their arms being designated as X1L, X1R, X2R, X2L, YL and YR. Meiotic behaviour of the sex trivalent was examined through the analysis of synaptonemal complexes (SCs), prometaphase I (metaphase I) and metaphase II nuclei. On the basis of the SC analysis, chromosomal length measurements at mitosis and prometaphase I and data from several orthopteran species, it is proposed that the breakpoints of the reciprocal translocation that originated this complex sex-determining mechanism were close to the centromeres of the X and the largest autosome, and that the asynapsed X1L and X2R regions observed in the sex trivalent at pachytene represent the original X chromosome. The X centromere being probably that of the X2 element because it lacks a partner in the SC pachytene trivalent. The relationship among synaptic pattern, chiasma localization and balanced segregation of the sex trivalent is also discussed.

Tension-sensitive kinetochore phosphorylation and the chromosome distribution checkpoint in praying mantid spermatocytes.

Improper chromosome attachment to the spindle can lead to daughter cells with missing or extra chromosomes. Such mishaps are avoided in many cells by a checkpoint that detects even a single improperly attached chromosome. What is detected? A misattached chromosome is not under tension from opposed mitotic forces, and in praying mantid spermatocytes, direct experiments show that the absence of tension is what the checkpoint detects. How is the absence of tension detected? Tension-sensitive kinetochore protein phosphorylation is the most likely possibility. We combined micromanipulation with immunostaining for phosphoproteins in order to study the effect of tension on kinetochore phosphorylation in mantid spermatocytes. We confirm earlier observations on mammalian cells and grasshopper spermatocytes that misattached chromosomes have phosphorylated kinetochore proteins. We also confirm experiments in grasshopper spermatocytes showing that tension alters kinetochore chemistry: tension from a micromanipulation needle causes kinetochore protein dephosphorylation, and relaxation of tension causes kinetochore protein rephosphorylation. Beyond confirmation, our results provide fresh evidence for phosphorylation as the signal to the checkpoint. First, mantid cells are the only ones in which an effect of tension on the checkpoint has been directly demonstrated; by equally direct experiments, we now show that tension affects kinetochore phosphorylation in these same cells. Second, sex chromosome behavior in mantids provides a natural experiment to test the relationship between phosphorylation and the checkpoint. In grasshoppers, an unpaired sex chromosome is normal, its kinetochore is under-phosphorylated, and the checkpoint is not activated. In mantids, exactly the opposite is true: an unpaired sex chromosome is abnormal, its kinetochore is phosphorylated and, as predicted, the checkpoint is activated. We conclude that tension-sensitive kinetochore protein phosphorylation very likely is the essential link between proper chromosome attachment and the check-point, the link that permits potential errors in chromosome distribution to be detected and avoided.

Neurons projecting from the brain to the corpora allata in orthopteroid insects: anatomy and physiology.

Retrograde and orthograde labeling of neurons projecting to the corpus allatum was performed in locust, grasshopper, cricket, and cockroach species in order to identify brain neurons that may be involved in the regulation of juvenile hormone production. In the acridid grasshopper Gomphocerus rufus L., and the locusts Locusta migratoria (R.&F.) and Schistocerca gregaria Forskal, the corpora allata are innervated by two morphologically distinguishable types of brain neurons. One group of 9-13 neurons (depending on species) with somata in the pars lateralis extend axons via the nervus corporis cardiaci 2 and nervus corporis allati 1 to the ipsilateral corpus allatum, whereas two cells in each pars lateralis have bilateral projections and innervate both glands. No direct connection between the pars intercerebralis and corpus allatum has been found. In contrast, neurons with paired axons innervating both glands are not present in Periplaneta americana (L.) and Gryllus bimaculatus de Geer. Instead, two cells in each pars lateralis project only to the gland contralateral to their somata. Electrophysiological experiments on acridid grasshoppers have confirmed the existence of a direct conduction pathway between the two glands via the paired axons of four cells that have been identified by neuroanatomy. These cells are not spontaneously active under experimental conditions. Ongoing discharges in the left and right nerves are unrelated, suggesting that the corpora allata receive independent neuronal inputs from the brain.

The midline metathoracic ear of the praying mantis, Mantis religiosa.

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