We investigated physical and chemical cues involved in male mating behavior of the white grub beetle, Dasylepida ishigakiensis (Scarabaeidae). When presented with female attractant pheromone (R)-2-butanol lures in a flight tunnel, nearly all males exhibited orientation and touching behaviors to freshly killed males and females and to intact glass models. Males landed and bent their abdomens on male and female bodies, but not on intact glass models. When treated with one female equivalent (FE) extract, washed immature male bodies and glass models both evoked stronger male responses than untreated equivalents, with the former eliciting a greater response than the treated glass models. Male responses to target male and female bodies decreased with increased numbers of washings of target bodies with organic solvents. These results suggest that the chemical factors that elicit male abdominal bending behavior are present on the body surface in both sexes. Washed immature male bodies treated with 1 FE or one male equivalent (ME) of extract induced strong male abdominal bending behavior. Washed mature female bodies treated with 1 ME extract also evoked male responses. Extracts of both sexes included factors eliciting male abdominal bending behavior. These results suggest that both physical and chemical cues derived from conspecifics cooperate to facilitate male mating recognition in D. ishigakiensis. The mating process of this species in the field is highly synchronized. Thus, after orienting to a female-like object, the only information males require by touching is whether the sex attractant pheromone that attracted them is indeed from a conspecific.

We investigated the effects of visual information on wind-evoked escape behavior in the cricket, Gryllus bimaculatus. Most agitated crickets were found to retreat into a shelter made of cardboard installed in the test arena within a short time. As this behavior was thought to be a type of escape, we confirmed how a visual image of a shelter affected wind-evoked escape behavior. Irrespective of the brightness of the visual background (black or white) or the absence or presence of a shelter, escape jumps were oriented almost 180° opposite to the source of the air puff stimulus. Therefore, the direction of wind-evoked escape depends solely depended on the direction of the stimulus air puff. In contrast, the turning direction of the crickets during the escape was affected by the position of the visual image of the shelter. During the wind-evoked escape jump, most crickets turned in the direction in which a shelter was presented. This behavioral nature is presumably necessary for crickets to retreat into a shelter within a short time after their escape jump.

Establishment of the anterior-posterior axis is an important event in the development of bilateral animals. A homeodomain transcription factor, Otx, is important for the formation of the anterior part of the embryo, and its mRNA is expressed in a continuous manner in a wide range of animals. This pattern of expression is thought to be important for the formation of anterior neural structures, but the mechanism that regulates Otx expression remains largely unknown. Towards understanding how the transcription of Otx is maintained in the cells of anterior neural structure, the sensory vesicle, during embryogenesis, we examined transcription regulatory mechanisms of Otx, using embryos of the ascidian, Ciona intestinalis, from the gastrula to tailbud stages, which have not been studied previously. We identified two genomic regions capable of mimicking the Otx expression pattern from the gastrula to tailbud stages. Putative transcription factor binding sites required for this activity were identified. Notably, distinct sets of transcription factor binding sites were required at different developmental stages for the expression of Otx, suggesting that the continuity of Otx is supported by distinct transcriptional mechanisms in the gastrula and neurula stages. Along with previous studies using Halocynthia roretzi, the present results provide insight into the evolution of transcriptional regulatory mechanism of Otx.

Eusocial insects have highly sophisticated societies, showing a conspicuous division of labor associated with different phenotypes. These castes show specific morphologies adapted to discrete tasks. Termite castes are divided into reproductives, workers, and soldiers. Individuals with soldier-like heads as well as developed gonads have been recorded in several primitive families, and are called reproductive soldiers. In some termite species, however, a trade-off-like developmental relationship has been shown between soldier and imaginal characteristics. Thus, while the mechanism that regulates the development of both characteristics in the same individual is interesting, the details are still unclear. We focused on juvenile hormone (JH), which is involved not only in termite caste differentiation, but also in the gonad development of many insects, and we aimed to clarify the effects of JH on the differentiation of reproductive soldiers in Zootermopsis nevadensis. We succeeded in the induction of individuals with reproductive soldier-like gross morphologies by JH analog (JHA) application to several developmental stages. We also observed that gonad development was significantly promoted by JHA application after molts in the induced reproductive soldier-like individuals, but not in natural soldiers. Finally, we confirmed that the gene expression level of vitellogenin was extremely high in the induced reproductive soldier-like individuals following JHA treatment after the molt. These results suggested that soldiers do not have regulatory mechanisms for gonad development involving JH, and the regulation of reproductive soldiers development is different from that of soldiers. Reproductive soldiers may have evolved independently from the soldier caste rather than from an intermediate stage of soldier evolution.

Myrmecina nipponica Wheeler is a terrestrial ant nesting chiefly in the soil in forest. It is a specialized predator of oribatid mites, but also scavenges on a broad spectrum of other arthropods. In the studied population at Cape Manazuru in central Japan, M. nipponica colonies are typically monogynous, and previous dissections of queens suggested that these individuals were not inseminated, thus suggesting these ants can reproduce via thelytokous parthenogenesis. To test for thelytokous parthenogenesis in M. nipponica the spermathecae of queens (dealate gynes) from worker-containing colonies were histologically examined in detail. All specimens examined (n = 5) had no spermatozoa in the spermatheca. In addition, a total of four colony-founding queens were reared in isolation in the laboratory to test whether non-inseminated females were capable of egg laying and to test whether female offspring emerged from this brood. In all of four culture replicates, only new workers were produced from the eggs those queens had laid and male offspring was absent. After the breeding experiment, the queens' spermathecae were histologically examined and no sperm were detected in their spermathecae. These results reveal that M. nipponica queens of the Manazuru population are capable of producing female offspring thelytokously. Sexual reproduction by typical gynes and also by intermorphs has been known from other local populations of M. nipponica; therefore, this species shows geographical polymorphism in sexuality.

Ground beetles of the subgenus Ohomopterus (genus Carabus) show marked divergence in species-specific male and female genital morphologies, which contributes to reproductive isolation among species. Characterizing the genetic basis of species-specific genital morphology is essential for understanding their diversification, but genomic information on Ohomopterus is not yet available. We analyzed mRNA extracted from abdominal sections of the last instar larvae and pupae of two sister species, Carabus (Ohomopterus) iwawakianus and C. (O.) uenoi, which show marked differences in genital morphology, to compare transcriptomic profiles using Roche 454 pyrosequencing. We obtained 1,608,572 high-quality reads and assembled them into 176,278 unique sequences, of which 66,049 sequences were combined into 12,662 clusters. Differential expression analyses for sexed pupae suggested that four and five clusters were differentially expressed between species for males and females, respectively. We also identified orthologous sequences of genes involved in genital development in Drosophila, which potentially affect genital development and species-specific genital morphology in Ohomopterus. This study provides the first large transcriptomic data set for a morphologically diversified beetle group, which can facilitate future studies on the genetic basis of species-specific genitalia.

Freshwater gastropods often have limited dispersal capability and small geographic ranges, and face severe threats from habitat loss and degradation. However, in addition to the scarcity of knowledge on their life history traits, species taxonomy has not been adequately resolved and boundaries between intra- and interspecific variation remain unclear for many taxa. One such example of an indeterminate species boundary with implications for conservation issues is the relationship between the thiarid snails Stenomelania crenulata in Okinawa and southwards (ranked as CR EN in the 2012 Japanese Red List) and S. rufescens in mainland Japan (VU). The results of our multi-disciplinary investigation into variation in the shell morphology and mitochondrial (COI) and nuclear (ITS-1) gene sequences suggest that S. rufescens represents a geographic variant and a junior synonym of S. crenulata. The widespread geographic range of S. crenulata, spanning a few thousand kilometers north to south, is possible due to an amphidromous life cycle that involves a marine planktotrophic larval phase and upstream migration after settlement in estuaries. Nevertheless, there is recognizable morphological and genetic differentiation between distant populations, probably reflecting a relatively short pelagic duration and possibly also infrequent transoceanic dispersal; metamorphic competence is achieved in two weeks in full seawater and even more rapidly in brackish water. The Okinawan population, with only a few known localities, therefore deserves the high conservation priority; conservation efforts need to involve the proper maintenance of migration pathways including all marine, brackish and freshwater environments.

Buthid scorpions exhibit a high variability in diploid number within genera and even within species. Cytogenetically, Buthidae differs from other families of Scorpiones based on its low diploid numbers, holocentric chromosomes, and complex chromosomal chains, which form during meiosis. In this study, we analyzed the distribution of the 45S ribosomal DNA (rDNA) genes in the mitotic and meiotic chromosomes of seven buthid species belonging to the genera Rhopalurus and Tityus with the ultimate goal of elucidating the chromosome organization in these scorpions. The chromosome number ranged from 2n = 6 to 2n = 28. Despite the high variance in diploid number, all species examined carried their 45S rDNA sites in the terminal region of exactly two chromosomes. Analyses of meiotic cells revealed 45S rDNA clusters in the chromosomal chains of Rhopalurus agamemnon, Tityus bahiensis, Tityus confluens, and Tityus martinpaechi, or in bivalent-like configuration in Rhopalurus rochai, Tityus bahiensis, Tityus confluens, Tityus fasciolatus, and Tityus paraguayensis. In the species examined, the 45S rDNA sites colocalized with constitutive heterochromatin regions. In light of the high chromosome variability and maintenance of number and terminal position of 45S rDNA sites in buthids, the heterochromatin may act to conserve the integrity of the ribosomal genes.

A vigorous discussion surrounds the question as to what enables some mammals—including primates and cetaceans—to evolve large brains. We recently published a study suggesting that the radiation of marsupial mammals is highly relevant to this question because of the unique reproductive and metabolic traits within this clade. In particular, we controversially suggested that marsupial brain sizes are not systematically smaller than those of placentals, and that elevated basal metabolic rates (BMR) are not linked to larger marsupial brains. As our dataset was found to contain some erroneous body size data, derived from a published source, we here use an updated and corrected dataset and employ standard as well as phylogenetically corrected analyses to re-assess and elaborate on our original conclusions. Our proposal that marsupials are not systematically smaller-brained than placentals remains supported, particularly when the unusually large-brained placental clade, Primates, is excluded. Use of the new dataset not only confirms that high metabolic rates are not associated with larger brain size in marsupials, but we additionally find some support for a striking negative correlation between BMR and brain size. The best supported correlates of large brain size remain the reproductive traits of weaning age and litter size. These results support our suggestion that mammalian brain sizes (including, by inference, those of monotremes) are predominantly constrained by the ability of females to fuel the growth of their offspring's large brains, rather than by the maintenance requirements of the adult brain.

Because all available evidence from allozymes, mtDNA sequences, and artificial hybridization suggests presence of high genetic differentiation between populations of East Asian brown frogs currently assigned to Rana dybowskii Günther, 1876, I compared morphological characters between specimens from Tsushima Island of Japan and Maritime territory of Russia. The population from Tsushima is slightly, but significantly different from R. dybowskii from Russia, including the holotype. I therefore consider the Tsushima population to be specifically distinct, and describe it as a new species R. uenoi. The new species also occurs in the Korean Peninsula and adjacent islands, but the distributional relationships with R. dybowskii are unclear, as detailed distribution in northern Korea is lacking.