Ogerin is a positive allosteric modulator of human and mouse ovarian cancer G protein-coupled receptors (OGR1s). In the present study, we found that ogerin differentially enhances the activation of OGR1 in various animal species. Amino acid residues of OGR1 that are associated with ogerin are conserved among the species. This suggests that other amino acid residues may be involved in the action of ogerin. Chimeric receptors between human and zebrafish OGR1s showed that the amino acid residues that determine the species specificity of ogerin-induced enhancement reside in the transmembrane and/or intracellular regions of OGR1. This result highlights the importance of first verifying the effectiveness of ogerin to the OGR1 of the species of interest at the cellular level prior to analyzing the physiological and pathophysiological roles of OGR1 in the species.

Genetic structures of two closely related butterflies, Ypthima multistriata and Y. argus, inhabiting Japan were compared based on the mitochondrial cytochrome c oxidase subunit I (COI) sequences. The former species is classified as a vulnerable species and exhibits a characteristic pattern of voltinism: univoltine and bivoltine populations are distributed in a scattered manner. The latter species is common and has a normal geographical pattern of voltinism: the number of annual generations is correlated with latitude. Our genetic analyses of these two species yielded contrasting results: a spatial analysis of molecular variance (SAMOVA) and F_ST between each pair of populations revealed a locally fragmented genetic structure for Y. multistriata, compared to three distinct geographic groups of Y. argus within which range-wide gene flow occurs. Although Y. argus is a common species, only the southernmost populations in Japan had higher genetic diversity, while the other populations had the same or lower levels of genetic diversity, compared to Y. multistriata. These results indicate that: 1) the degree of fragmentation of Y. multistriata populations was higher; however, markedly lower genetic diversity was not found, and 2) although Y. argus is a common species, its populations may not be genetically robust. In addition, AMOVA revealed a relationship between voltinism and genetic variation in Y. multistriata. This result suggests a phylogenetic constraint of voltinism in this butterfly.

The parasitoid wasp Protaphidius nawaii parasitizes the aphid Stomaphis japonica, which is obligatorily attended by several species of ants of genus Lasius. Subgenus Lasius or Dendrolasius ants use different defense strategies to protect the aphids that they attend (Lasius, shelter building; Dendrolasius, aggressive attack). We performed molecular phylogenetic analysis based on partial mitochondrial DNA sequences of P. nawaii and found that the parasitoid wasp consists of two highly differentiated genetic lineages. Although these two lineages distributed sympatrically, one tends to parasitize aphids attended by ants of subgenus Lasius, and the other parasitizes aphids attended by ants of subgenus Dendrolasius. The two lineages of P. nawaii appear to exhibit different oviposition behaviors adapted to the different aphid-protection strategies of the two ant subgenera.

Pseudogenization of the umami taste receptor gene Tas1r1 has been reported more commonly in aquatic than terrestrial mammals. We hypothesized that the more species are adapted to the aquatic environment, the less important a role the umami taste detection plays. To test this hypothesis, we focused on the Mustelidae family because their habitat and feeding ecology are highly diverse. We found pseudogenizing mutations in exon 1 of Eurasian otter and exon 6 of African clawless otter, both of which forage underwater. Our analysis of the ratio of nonsynonymous to synonymous nucleotide substitution rates suggested that purifying selection pressures on Tas1r1 are weaker in the lineages with non-functional Tas1r1 than the lineages retaining functional Tas1r1. Our analysis also suggested that relaxed selection pressures on Tas1r1 in Mustelidae species adapted to the aquatic environment, although we cannot exclude the possibility that they are restricted to Lutrinae irrespective of their feeding habitat. Overall, the results of the present study support the idea that differences in selection pressures on Tas1r1 reflect differences in feeding behaviors.

The family Sicariidae comprises the genera Hexophthalma, Sicarius and Loxosceles. This latter is subdivided in eight monophyletic groups based on genitalia morphology and molecular analyses: amazonica, gaucho, laeta, and spadicea (South America); reclusa (North America); rufescens (Mediterranean); spinulosa and vonwredei (Africa). In Brazil, the genus Loxosceles is represented by 50 species. The mitotic and meiotic characteristics of eight Loxosceles species were analyzed in order to discuss the chromosome evolution, as well as the correspondence between cytogenetic data and morphological/molecular data for the delimitation of the South American groups of species belonging to this genus. All species studied in this work showed 2n♂ = 23, including a X1X2Y sex chromosome system (SCS). Despite the similarity of diploid number and SCS, the species studied here differed regarding the chromosome morphology of some autosomal pairs, presence of secondary constrictions, size of X chromosomes and localization of Ag-NOR/rDNA sites. Based on all these chromosomal data, we verified a close relationship between Loxosceles species belonging to the amazonica and gaucho groups. Transmission electron microscopy (TEM) analysis of spread pachytene cells of L. gaucho showed regular synapsis between homologous autosomal chromosomes, but asynaptic behavior of the sex chromosomes. The axial elements of the sex chromosomes undergo conspicuous morphological modifications resulting in shortening of their length.

To elucidate the role of dmrt1 in sex differentiation of a teleost fish Schizothorax kozlovi, the full-length sequences of its cDNA and promoter were cloned by rapid amplification of cDNA ends (RACE) and genome walking. The relative mRNA expression levels were determined by quantitative real-time PCR (RT-PCR). The 1095-bp dmrt1 cDNA was predicted to encode a protein of 264 amino acids. It was expressed only in the gonads, and the expression was 17-times higher in the testis than in the ovary. The 1215-bp promoter sequence of dmrt1 was cloned and analyzed to detect sex-related differences in its methylation levels. A significant negative relationship between the dmrt1 expression and CpG methylation of its promoter were found in the testes and ovaries of S. kozlovi. Significant differences in dmrt1 expression levels were also found between the larval and juvenile stages. No significant differences in expression were found during the entire larval stage, and in the individuals among three different temperature groups (10°C, 14°C, and 18°C). Considering that the sex of sampled larval fish cannot be distinguished, correlations between dmrt1 expression and effects of temperature on sex differentiation in S. kozlovi need further study.

In the present study, we observed that Leptinaria unilamellata responds to changes in microclimatic conditions through shifts in shell morphology. Over three laboratory generations, shell differences between two populations, representing distinct morphotypes, became less evident. Only F1 generations from both populations showed shell morphometry very similar to the field parental snails, suggesting maternal effect. Snails from the locality with higher values of rainfall, relative humidity and evaporation index and smaller values of temperature and insolation produced more and larger hatchlings. Snails from the locality with less favorable climatic conditions presented shells traits that offer protection against desiccation, but reduce reproductive success. These snails showed smaller offspring production and faster response to a desiccation regime, through changes in conchiometrics. In addition, the results of the present study suggest that the spire index plays a less important role in determining protective properties of the shell of L. unilamellata, in response to desiccation risk, compared to aperture dimension. As shell aperture dimension is an important trait related to resistance to desiccation, and at the same time to reproduction, plastic responses to environmental conditions promoting the balance between survival and reproductive success are critical for the species adaptive success.

Kawai et al. (2011) recently introduced a mixture of three anesthetic agents (here called MMB) that has an effect similar to ketamine/xylazine in mice, which might allow more effective reaction to changes in the animal condition, as an antagonist is available, and which can be used without license for handling narcotic drugs. Using Kawai's study as a baseline, we tested whether this anesthesia and its antagonist can also be used in avian studies. In the present study, we used two species, the zebra finch and the Bengalese finch, of the avian family Estrildidae. In zebra finches, anesthesia effects similar to the use of ketamine/xylazine and to those obtained in mice can be reached by the use of MMB if a higher dose is applied. MMB leads to more variable anesthesia, but has the advantage of a longer time window of deep anesthesia. An antagonist to one component of MMB reduced the awaking time, but was not as effective as in mice. For Bengalese finches, MMB cannot be generally recommended because of difficult handling and high mortality rate when used without antagonist, but could be used for perfusions instead of pentobarbital.

In terrestrial vertebrates, the hippocampus plays a major role in spatial cognition. Recent developmental, anatomical, and histological studies suggest that the ventral region of the lateral part of the dorsal telencephalic area (Dlv) in teleost fishes is homologous to the hippocampus in terrestrial vertebrates. We hypothesized that fish species with higher spatial cognitive demands have a more highly developed Dlv compared to closely related species with relatively lower spatial cognitive demands. The fishes selected for this study were Favonigobius gymnauchen, Istigobius hoshinonis, Tridentiger trigonocephalus, and Chaenogobius annularis; all are bottom-dwelling gobiid species found in habitats that vary with respect to their spatial complexity. Volumetric analysis of the telencephalic subregions, including the Dlv, and other major brain regions showed that species from stable rocky areas had a larger Dlv than species from relatively homogenous sandy/ muddy environments. These findings support the hypothesis that the teleost Dlv is homologous to the hippocampus in terrestrial vertebrates, and that the relative development of these areas is positively correlated with spatial cognitive demand in animals.

There are few model fish that are both edible and suitable for use in the laboratory. The Japanese loach (Misgurnus anguillicaudatus) is a traditional food in Japan, but is highly neglected despite its great nutritional value. To understand its circadian system and photic input pathway for synchronization of physiological activities to environmental light-dark cycles, we measured locomotor activity under light-dark and constant dark (DD) conditions. Locomotor activity was found to be higher in the nighttime than daytime, and its rhythmicity was weakened under DD conditions. The nocturnal activity of the Japanese loach is mainly controlled by environmental light, rather than the circadian clock. We explored the circadian regulation and light-responsiveness of clock gene expression in the eyes of loaches. The daily expression profiles of its mRNA revealed that most of the examined Cry and Per genes were likely regulated by internal circadian and/or environmental light signals. Among the Opsin genes transcribed in the eye, we detected the retinal photopigment porphyropsin at the protein level, which was lower than in mice. This property of loach eyes prompted us to analyze the locomotor activities of eye-enucleated fish. As a result, they still showed nocturnal circadian activity. Thus, it is likely that extraocular photoreceptive tissue(s) also contribute to the photic input pathway, although loach eyes are a circadian photosensitive tissue. This suggests that the loach mainly uses not its vision but other stimuli, such as mechanical or chemical stimuli, detected by barbels, to coordinate its nocturnal behavior.

The Japanese wrinkled frog Glandirana rugosa is separated into five genetically different groups. One group in western Japan is further divided into three subgroups, found in Kyushu, Shikoku, and western Honshu. We collected G. rugosa frogs at 39 sites in Kyushu and determined nucleotide sequences of the mitochondrial 12S and 16S rRNA genes for phylogenetic analysis. Unexpectedly, we found a group of frogs in southeastern Kyushu that did not cluster with any of the pre-existing five groups of G. rugosa on the phylogenetic trees. The frogs in the new group and G. rugosa in Kyushu were externally similar, but there were a few significant differences in morphological features between the two populations. In addition, we observed significant differences in the frogs' calls . Thus, the group of the frogs in southeastern Kyushu may represent a new candidate species in the genus Glandirana. We discuss the possibility of a new species.