Zoological Science

In this study, we investigated the chemotactic response of a wild-type (N2) nematode (Caenorhabditis elegans) to a water-soluble attractant, sodium acetate, after pre-exposure to the chemical. The chemotactic response to 1.0 M sodium acetate of the non-exposed control nematodes was lower than that of the nematodes that were pre-exposed to 1.0 M sodium acetate for 90 min (p < 0.05). The increase in the response to sodium acetate was observed up to 6 hr, but not at 12 hr after exposure. To clarify the mechanism of this enhancement of the chemotactic response, several mutants were used. The chemotactic response of pre-exposed tph-1 and bas-1 mutants, whose main defect was serotonin secretion, was enhanced in comparison with that of the control mutants (p < 0.01). However, cat-1 and cat-2 mutants, which are respectively defective in serotonin and dopamine secretion and dopamine secretion only, showed no enhancement of the chemotactic response to sodium acetate, even when pre-exposed to this chemical. When the cat-1 and cat-2 mutants were pre-exposed to sodium acetate and bred in the presence of 40 mM dopamine, these mutants showed enhanced chemotactic response to sodium acetate (p < 0.05). These results suggest that the enhancement of chemotactic response to sodium acetate after pre-exposure to this chemical is modulated by dopaminergic neurotransmission.

The large micromeres of the 32-cell stage of sea urchin embryos are autonomously specified and differentiate into primary mesenchyme cells (PMCs), giving rise to the skeletogenic cells. We previously demonstrated that HpEts, an ets-related transcription factor, plays an essential role in the specification of PMCs in sea urchin embryos. In order to clarify the function of HpEts in the gene regulatory network involved in PMC specification, we analyzed the zygotic expression pattern and the cis-regulatory region of HpEts, and examined the activity of the HpEts protein as a transcription factor. Intron-based PCR reveals that zygotic expression of HpEts starts at the cleavage stage, and that the rate of transcription reaches maximum at the unhatched blastula stage. A series of progressive deletions of the fragments from -4.2 kbp to +1206 bp of the HpEts, which directs PMC-specific expression, caused a gradual decrease in the specificity, implying that coordination of several cis-regulatory elements regulates the expression in PMCs. A minimum cis-element required for the temporal expression is located within a 10 bp from -243 bp to -234 bp. The HpEts protein remains in the cytoplasm of entire embryonic cells in the cleavage stage. At the unhatched blastula stage, the HpEts protein translocates into the nucleus in presumptive PMCs. Transactivation assays demonstrate that the HpEts protein activates a promoter of Spicule Matrix Protein 50 (SM50), which is a target of HpEts, which binds to the regulatory region of SM50.

Mitochondrial and nuclear DNA analyses were used to examine the native distribution range and population structure of Biwia zezera (Cyprinidae: Gobioninae) based on specimens from the species' presumed distribution range. We found two greatly differentiated groups, with 8.6% uncorrected sequence differences in the mtDNA cytochrome b gene; one group was distributed exclusively in the Yodo River system (excluding Lake Biwa and rivers flowing into the lake). This differentiation was supported by results from three nuclear DNA loci, and it was concluded that the Yodo population is an undescribed cryptic species. Based on mtDNA haplotype distribution and endemicity, B. zezera was determined to be indigenous to the Ise Bay area (Nobi Plain), Lake Biwa basin (excluding the Yodo River outlet and its tributaries), the Sanyo region, and northern Kyushu. The population in the Ise Bay area was the most differentiated from the others, while that in Kyushu was the second-most differentiated. It was confirmed that an endemic population closely related to the Lake Biwa population is naturally distributed in the Sanyo region.

We examined whether gonadotropin-releasing hormone (GnRH)-like peptides are present in the neural ganglia of the gastropod Pacific abalone (Haliotis discus hannai) by reverse-phase high performance liquid chromatography (rpHPLC) combined with time-resolved fluoroimmunoassay (TR-FIA) analysis and by immunohistochemistry. Cerebral ganglion extracts showed a similar retention time to lamprey GnRH-II (lGnRH-II) in rpHPLC combined with TR-FIA analysis. GnRH-like-immunoreactive (ir) cell bodies (which reacted with a mouse monoclonal antibody raised against the common amino acid sequence of vertebrate GnRH) were detected in the peripheral region of the cerebral ganglion, and they were observed to send fibers into the neuropil. GnRH-like-ir fibers were also detected in the neuropil of the pedal ganglion, the visceral nerve, and the nerve originating from the pedal ganglion. Chicken GnRH-II (cGnRH-II)-like-ir fibers (which reacted with a rabbit polyclonal antibody raised against cGnRH-II) were also observed in the neuropil of the cerebral ganglion. GnRH-like-ir fibers and cGnRH-II-like-ir fibers were distinguishable in the neuropil of the cerebral ganglion by double-staining immunohistochemistry. These results suggest that multiple GnRH-like peptides exist in the neural ganglia of the Pacific abalone.

The complete sequences of mitochondrial DNA (mtDNA) from the three new grasshopper species, Euchorthippus fusigeniculatus, Mekongiana xiangchengensis and Mekongiella xizangensis, consisting of 15772 bp, 15567 bp, and 15885 bp, respectively, were analyzed and compared to mtDNAs from other 19 Orthoptera species obtained from GenBank. The three mitochondrial genomes contain a standard set of 13 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes. and an A+T-rich region in the same order as those of the other analyzed caeliferan species, but different from those of the ensiferan species by the rearrangement of trnD and trnK. The putative initiation codon for the cox1 gene is ATC in E. fusigeniculatus, CTG in M. xiangchengensis and CCG in M. xizangensis. All secondary structures of tRNA-Ser^(AGN) in the three species lack a DHU arm. In this study, we stressed the comparative analysis of the stem-loop secondary structure in A+T-rich region of all Orthoptera species available to date, and report new findings which may facilitate further investigation and better understanding of this secondary structure. Finally, we undertook a phylogenetic study of all Orthoptera species available from GenBank to date based on three different datasets using parsimony, maximum likelihood, and Bayesian inference. Our result showed that protein-coding genes (PCG) and amino acid sequences (PCG_PROT) provided good resolution of higher-level relationships within the Orthoptera, whereas ribosomal RNA genes (RIBO) perform poorly under different optimality criteria.

The exoskeleton of arthropods undergoes reformation at every molting. Accordingly, external morphology can metamorphose through molting. In some crustaceans, the function of appendages is modified through ontogeny. These morphological modifications require accordant modification of the correlation between different body parts because the morphological function depends on the combined correlation between different parts. In the case of crustacean morphology, exoskeleton and muscles are correlated to each other. The functional morphology of the fifth limb of cypridoid ostracods transforms from “walking leg + mouthparts (+ possibly respiratory parts)” to “mouthparts + respiratory parts + grasping hook (in males only)” through ontogeny. In this study, the three-dimensional structures of the exoskeleton and muscular systems were observed by confocal laser-scanning microscopy in some species of suborder Cypridocopina. The muscular system is reportedly not changed by the ontogeny of appendages in females, but it does change in males. Furthermore, regional cell proliferation, which was detected previously, represented the causal factor of exoskeletal modification. I therefore conclude that the enlarged endite in the female fifth limb is produced by exoskeletal modification based on regional cell proliferation, rather than by a change in the muscular system. In contrast, modification in the male requires a change in the muscular system in addition to exoskeletal modification.

We describe a new species, Tegastes okinawaensis, from 583–711 meter-deep hydrothermal vents in the Okinawa Trough, Japan. This species is clearly distinguishable from its congeners by four combined characters, namely 1) the last three abdominal somites are depressed inwardly towards the body, 2) there is sexual dimorphism in the number of setae of P3 and P4, 3) the male antennule is 9-segmented, with the 7th segment formed by incomplete fusion of two segments, and 4) the caudal rami are small with a distorted quadrangular shape. Additionally, the species is easily distinguishable from its congeners by the distal endopodal segment of P4, which is armed with six setae — this character has never been reported in previous descriptions of congeners. This report is the first of a species in the genus Tegastes living at ocean depths of greater than 580 m in the northwestern Pacific.

A new genus and species of cyclopoid copepod belonging to the family Enterognathidae, Parenterognathus troglodytes, is described from a gall on the calyx of the deep-sea crinoid Glyptometra crassa (Clark, 1912) collected at depths of 775–787 m off Kumano-nada, middle Japan. The new genus can be distinguished from the three known genera of the family by body tagmosis and by the segmentation and armature of the appendages. This is the first record of this family from the Pacific Ocean. This family seems to be host-specific to relatively basal deuterostomes, such as echinoderms and hemichordates. The evolutionary transformation and history of the Enterognathidae are briefly discussed.

The genua Roquettea Mello-Leitão, hitherto monotypic and known from Brazilian state of Pará, is rediagnosed, the type species, Roquettea singularis is redescribed, including a report on two male morphs, being the second record of male polymorphism in Cosmetidae, the first in South America. Three new species are described from northern Brazil — Roquettea taurine n. sp. (which possesses a unique horned ocularium) and Roquettea jalapensis n. sp. (without notable scutal structures), both from the state of Tocantins, being the first record of the family Cosmetidae from Tocantins; and Roquettea scrotalis n. sp. (with a unique 2-balled dorsal tuberous complex) from Amapá state, being the first record of the subfamily Discosomaticinae from Amapá. Roquettea is compared to the closely related genus Gryne, both currently placed in Discosomaticinae, and also to Profus, the type genus of the subfamily. The unity of Discosomaticinae is discussed, and evidence for the monophyly of this subfamily is weak at best. Male genitalia are for the first time illustrated and described for genera Protus, Roquettea and Gryne, and SEM micrographs are used for the first time in descriptions of Cosmetidae.

Dual-color fluorescence in situ hybridization (FISH) was performed to study the simultaneous localization of major (45S) and minor (5S) family ribosomal RNA genes on chromosomes of Neolissochilus hexagonolepis. The partial 45S (18S, ITS 1, 5.8S, ITS 2 and 28S) and complete 5S (coding and NTS) rDNA units were amplified, sequenced, analyzed, and mapped on the metaphase chromosomes. The complete 18S, 5.8S and partial 28S rDNAs were 1849 bp, 157 bp and 1819 bp long, respectively. Internal transcribed spacers, namely ITS 1 (828 bp) and ITS 2 (359 bp), showed significant nucleotide variations from other fish species listed in NCBI database. The 5S rDNA contained an identical coding region of 120 bp and a highly divergent, non-transcribed 81-bp spacer. The specimens of N. hexagonolepis showed six bright fluorescent signals of 18S, while the 5S signals were present only on one pair of chromosomes. Subsequent analyses between conventional Ag-NORs and 18S rDNA FISH strongly suggested the possible inactivation of one pair of NORs that was localized at a telomeric position of a submetacentric chromosome. The sequencing and chromosomal localization of 45S and 5S rDNAs may serve as a useful genetic marker in taxonomic classification as well as in phylogenetic and evolutionary studies.