We assessed the historical biogeography of the Ryukyu five-lined skink, Plestiodon marginatus, and related species (P. stimpsonii and P. elegans). Our specific aims were to reveal the origin, tim- ing, and route of the colonization to three volcanic islands in the northern Tokara Group of the northern Ryukyus: Kuchinoshima, Nakanoshima, and Suwanosejima. We conducted phylogenetic analyses and divergence time estimation using a partial sequence of the mitochondrial cytochrome b gene for P. marginatus collected from across its whole range (the northern and central Ryukyus), and for P. stimpsonii (from the Yaeyama Group of the southern Ryukyus) and P. elegans (from Taiwan). Our results suggest three major clades (A, B, and C). Clades A and B consist of P. marginatus, excluding the Kuchinoshima population, and Clade C consisted of the Kuchinoshima population, P. stimpsonii, and P. elegans. These clades are estimated to have diverged during the Late Miocene to the Late Pliocene. Among the three examined northern Tokara populations, the Kuchinoshima population was shown to be a sister group of P. stimpsonii. The two other populations from Nakanoshima and Suwanosejima Islands were closely related to P. marginatus from the northern part of the Okinawa Group and that from Kodakarajima Island in the southern Tokara Group, respectively. These populations are estimated to have diverged from their respective related spe cies in various ages of the Early to Late Pleistocene, suggesting that they colonized the islands by independent overseas dispersals of approximately 50–850 km via the Kuroshio Current. Taxonomic implications for P. marginatus are also discussed.

The transactivational property of natural and synthetic chemicals via medaka vitamin D receptor β subtype (VDRβ) was investigated after the development of a stable cell line expressing a Gal4-VDRβ fusion protein for reporter gene assay. Members of vitamin D class, including 1α, 25- dihydroxyvitamin D₃ (1,25VD3) were specifically detected as agonists in our system. Although other steroids and chemicals used in the present estimation induced no agonistic response, 10 compounds displayed antagonistic or synergistic activity. Spironolactone, which is an antagonist of corticoid receptors in mammals, competitively inhibited the transactivity of 1,25VD3 by over 80% in a dosedependent manner. Mifepristone and cyproterone acetate were also detected as antagonists, but they significantly acted only at 10µ. Pregnenolone and raloxifene dose-dependently enhanced the activity of 1,25VD3 at EC₅₀ to the maximum level. Diethylstilbestrol, 17α-ethynylestradiol, genistein, and stanozolol were also synergists, but their potency was low. Interestingly, dibutyltin dichloride, which is used as a stabilizer in the production of polyvinyl chloride plastics, produced greater response than maximum effect of 1,25VD3 although the concentration-response curve was not typically sigmoidal. In the present study, we successfully developed a stable reporter gene assay, which allows assessment of the vitamin D-like chemicals toward the medaka VDRβ.

The gene, vitellogenin (vtg) was cloned and characterized in the dojo loach (Misgurnus anguillicaudatus), an indigenous freshwater species in East Asia, in order to develop tools for detecting the effects of estrogenic endocrine-disrupting chemicals (EEDCs). Full-length cDNAs encoding seven distinct vtg transcripts (vtg1–7) were obtained. The corresponding deduced amino acid sequences (Vtg1–7) were divided into two types; type I (Vtg1–6; 89–99% identical), which contained both lipovitellin (Lv) and phosvitin (Pv), and type II (Vtg7), which contained Lv alone. Phylogenetic analysis revealed that the type I and type II Vtgs in the loach could be classified as VtgAo1 and VtgC types, respectively. Immuno-biochemical analyses using type-specific Vtg antisera revealed that VtgAo1 proteins appeared to be the major Vtg type in this species. Males were administered (aqueous exposure) either 17β-estradiol (E2) or 17α-ethinylestradiol (EE2), the results from which were used to determine that hepatic vtgAo1 expression was estrogen-sensitive. The precise classification of the loach vtg/Vtg products, as well as their induction profiles following the estrogenic stimulation, provide a basis for their use as sensitive biomarkers when EEDC activities are evaluated in the freshwater environments in East Asia.

We investigated the salivary glands in Lepyronia coleopterata (L.), and found that the salivary glands are paired structures and consist of principal and accessory glands. Each principal gland contains an anterior lobe and a posterior lobe. Three types of acini (I, II, III) are observed in the anterior lobe, whereas the posterior lobe contains only one type of acini (IV). Rhabdus emerges from the middle portion of the acini III and IV. The oval-shaped accessory gland connects with the principal gland via a long duct. The long duct consists of a slightly coiled basal segment and a highly convoluted distal segment, with the terminal end of the latter constricted and connected with the accessory gland. A slightly convoluted transparent tube connects with the accessory gland at the former's distal end. The accessory gland, accessory salivary duct and the accessory salivary tube are observed for the first time in spittlebugs. Ultrastructurally, each type of acinus is made up of one type of secretory cells, but the rhabdus comprises two types of cells. Secretory granules in different type of cells are different in size, shape and electron density, which indicate either different materials are synthesized or these materials undergo a process of maturation. The rhabdus is empty in structure and contains several channels, with the lumen filled with abundant fine granular materials. Fine dark granules existed in the periphery of some secretory granules are probably virus particles. Microorganisms are observed in the cells of the acini I, III and rhabdus.

Associations among feeding habit, beak type, and food source in birds have been widely studied and are well known to exist. The relationship between feeding habit and jaw apparatus in birds has not attracted attention from ornithologists, perhaps because of the complexity of the skeletal morphology of the feeding system of birds. The goal of this study was to compare the jaw apparatus and foraging strategies of various Oriental species of the Picidae (Meiglyptini and Picini tribes) using a morphofunctional analysis of the skeletal structure of the jaw apparatus. This study showed that there are at least three types of jaw apparatus in these woodpeckers, as follows: 1) robust, developed, and complex; 2) complexity and development intermediate, as observed in Meiglyptes tristis and Dinopium spp., whose main foraging method involves gleaning, probing, and tapping; and 3) poorly developed, as observed in Picus miniaceus and Hemicircus concretus. The success of woodpeckers as a natural group is due not only to their feeding diversity, but also their ability to explore a wide range of different resources, as appropriate to their jaw apparatus.

Scincella boettgeri and S. formosensis are respectively small-bodied, morphologically similar skinks, endemic to the Southern Ryukyus and Taiwan. To estimate the phylogeography of both species, we performed phylogenetic analyses using the mitochondrial cytochrome b (cyt b) gene sequences based on 102 individuals of S. boettgeri from 12 Southern-Ryukyu islands and 33 S. formosensis from six localities in Taiwan. A total of 67 haplotypes were recognized for S. boettgeri and 21 for S. formosensis. The phylogenetic analyses revealed that the populations of Scincella spp. in the Southern Ryukyus and Taiwan are composed of three major clades, the Yonagunijima clade from Yonagunijima Island, the Southern-Ryukyu clade from the Southern Ryukyus exclusive of Yonagunijima Island, and the Taiwan clade from Taiwan. These clades showed high levels of genetic divergence, suggesting that the species have been isolated since the Early Pliocene. The Southern-Ryukyu and Taiwan clades were further divided into three and four subclades, respectively. Two of the three Southern-Ryukyu subclades are partially sympatric on two islets of the Yaeyama Group, suggesting this population represents a secondary contact subsequent to their allopatric differentiations.

The adult cricket Gryllus bimaculatus has a central clock in the optic lobe that regulates overt activity rhythms and secondary oscillators in the tissue outside the optic lobe. Here we investigated properties of the rhythmic expression of clock genes in the optic lobe and extra-optic lobe tissues in nymphs, and compared them with those of adults. In the optic lobe, mRNA of the clock genes period (per), timeless (tim), cycle (cyc) and Clock (Clk) were expressed in patterns similar to those in adult profiles, but at significantly lower levels. Among the extra-optic lobe tissues, the brain and TAG showed a rhythmic expression of per and tim, the mid-gut only in tim, and the anterior-stomach in none of the genes studied. The mRNA levels of clock genes were again significantly lower than those in adults. Unlike in adults, the brain and mid-gut lost their rhythms of clock gene expression in DD, and when the optic lobes were bilaterally removed. These results suggest that the rhythms outside the optic lobe are weak in nymphs, and may become robust after the imaginal molt.

A comparative analysis of reproductive effort, fecundity, and egg weight was conducted in two species of spider crabs, Leurocyclus tuberculosus and Libinia spinosa, during one-year period. Ovigerous females were collected from Patagonia-Argentina (42°56′S, 64°21′W) and were measured (CW = carapace width). Each egg brood was weighed, dried and the number of eggs (F = fecundity) counted. Scatterplots of relative fecundity (RF = F/CW) were submitted to regression analyses. Mean F and RF was calculated for each season to assess seasonal variation of reproductive intensity. Mean F was 35,000 eggs in L. tuberculosus and 30,000 eggs in L. spinosa, with these values being intermediate in comparison with other Majoidea. The RF was approximately 18% higher in L. tuberculosus that presented an average dry weight egg 45% less than L. spinosa. Although in both species F showed a positive correlation with CW, less than the 20% of the variation in the number of eggs could be explained by female's size, suggesting there are other factors that influence F. The proportion of body energy devoted to reproduction (reproductive effort), exhibited significant differences between species. In Leurocyclus tuberculosus reproductive activity is significantly different along the 12-month suggesting that the conditions for ‘optimal’ egg production change throughout the year. In Libinia spinosa mean fecundity did not reveal significant differences over seasons. These results are central in studies of life-history theory and in the development of lifehistory models, as it is directly related to energy allocation and partitioning.

Vitellogenesis has been extensively studied in oviparous vertebrates, including teleost fishes, while not much is known with regard to jawless hagfishes, modern representatives of the most primitive vertebrate class. This study aimed to characterize vitellogenin (Vtg) and yolk protein (YP) in the inshore hagfish (Eptatretus burgeri) as an initial step to understand vitellogenesis in this species. A putative Vtg fraction was purified from the serum of female hagfish by combinations of hydroxylapatite and ion-exchange chromatography, followed by gel filtration. The purified fraction appeared to contain two distinct Vtgs (Vtg1 and Vtg2) and exhibited biochemical properties resembling those previously reported for teleost Vtgs; these appeared to be female-specific serum proteins and high-molecular-weight proteins in gel filtration (˜505 kDa as the mixture fraction of both Vtgs) and in SDS-PAGE (Vtg1 and Vtg2; ˜210 kDa and ˜195 kDa, respectively). A major YP was also purified from hagfish eggs by combinations of hydroxylapatite chromatography and gel filtration; the apparent native mass of the purified YP was unusually large (> 669 kDa). The purified YP consisted of four polypeptides in SDS-PAGE; the peptide pattern indicated that it consisted of two lipovitellins (Lv1 and Lv2) giving rise to two sets of heavy chains (˜116 kDa and ˜106 kDa, respectively) and two light chains (˜32 kDa and ˜28 kDa, respectively). Additional immunological analysis, Nterminal amino acid sequencing and cDNA cloning firmly confirmed the precursor-product relationship between hagfish Vtgs and Lvs.

Wockia, one of the six genera within the lepidopteran family Urodidae, currently includes nine species distributed in Holarctic, Oriental, and Neotropical regions. A new species of the genus, W. magna sp. nov., is described from Japan and South Korea. This is the first record of Urodidae from Japan. A cladistic analysis was constructed based on 27 morphological characters from five ingroup species and two outgroup taxa. A single most parsimonious tree was found (length = 38, CI = 71, RI = 70). The resulting tree failed to recover the monophyly of Wockia chewbacca and a clade Wockia sensu stricto, including four congeners from North America, temperate East Asia and Southeast Asia. Wockia sensu stricto was resolved as monophyletic and was divided into two subgroups, one including W. asperipunctella and W. magna, and the other including W. koreana and W. variata. The sister-group relationship of W. asperipunctella and W. magna was moderately supported. Synapomorphies of Wockia sensu stricto are provided from the resulting phylogeny. Systematic definitions of Wockia and other allied genera are revised. Optimal ancestral area reconstruction implemented in DIVA resulted in different hypotheses for Wockia, depending on how to constrain the maximum number of areas. The maximum area number set as two yielded a more likely scenario suggesting that Wockia sensu stricto originated in temperate East Asia and then dispersed into the Oriental region, North America, and Europe. A working hypothesis and other equally possible alternative explanations for the biogeography of Wockia are provided.