Ovulation is the process of liberating oocytes from the preovulatory follicles, and is observed in the ovaries of virtually all female vertebrate animals. Compared with mammalian species, there have been far fewer studies that address the ovulatory mechanisms of non-mammalian species. We have examined the molecular mechanism of follicle rupture during ovulation using the teleost model, medaka, or Oryzias latipes. Follicle rupture in medaka ovulation involves the cooperation of the tissue inhibitor of metalloproteinase-2b protein with at least three matrix metalloproteinases (MMP): membrane type-1 MMP (MT1-MMP), MT2-MMP, and gelatinase A. Our studies also indicate that the serine protease, i.e., plasmin, participates in the rupture for only a few hours prior to the activation of MMP-mediated hydrolysis at ovulation. The involvement of prostaglandin E₂ (PGE₂) in medaka ovulation was also demonstrated. Cyclooxygenase-2 and PGE₂ receptor subtype EP4b were respectively shown to be an enzyme responsible for PGE₂ synthesis and a receptor for the generated ligand in the preovulatory follicles. Based on the results obtained from our studies of fish, we discuss the similarities and differences in vertebrate ovulation compared with mammalian species.

Rodent scent-marking is often used for territorial defence and self-advertisement, and both functions often entail the continuous scent-marking of a large area with high costs. In species with highly-fragmented populations and low density, in which the likelihood of social encounters is low, the costs of continuous scent-marking might exceed the associated fitness benefits; therefore, less intensive scent-marking only to signal presence to the opposite sex may be used. This hypothesis was tested in captivity with the Cabrera vole, a species with highly fragmented and low-density populations. Firstly, to assess the unknown scent-marking behaviour of the Cabrera voles, we conducted an assay wherein voles could scent-mark a clean substrate. Both sexes marked with urine and faeces, but never with anogenital secretions, and the amount of scent-marks was not different between sexes. In the subsequent assay, voles of each sex were given the choice of scent-mark on clean substrates or on substrates previously scent-marked by males or females. Both sexes marked with urine a larger area on substrates pre-marked by the opposite sex than on substrates pre-marked by the same-sex and clean substrates; however, no differences were found in the frequency of fecal boli deposited on the three types of substrate, and no anogenital secretions were found. The clear preference of receivers to scent-mark with urine the substrate pre-marked by the opposite sex strongly suggests that Cabrera voles use urine scent-marking for inter-sexual communication, probably to increase mate-finding likelihood, rather than for territorial defense and/or self-advertisement.

The time-honored assumption about the defensive function of trichocysts in Paramecium against predators was recently verified experimentally against different species of unicellular predators. In the present study, we examined the defensive function of trichocysts against three metazoan predators, Cephalodella sp. (Rotifera), Eucypris sp. (Arthropoda), and Stenostomum sphagnetorum (Platyhelminthes). The results confirmed the defensive function of trichocysts against two of these metazoan predators (Cephalodella sp. and Eucypris sp.), while they seem ineffective against S. sphagnetorum. We also compared the defensive efficiency of the trichocysts of P. tetraurelia with that of toxin-containing extrusomes of two ciliates.

Various selection pressures induce the degree and direction of sexual size dimorphism in animals. Selection favors either larger males for contests over mates or resources, or smaller males are favored for maneuverability; whereas larger females are favored for higher fecundity, or smaller females for earlier maturation for reproduction. In the genus of Larus (seagulls), adult males are generally known to be larger in size than adult females. However, the ontogeny of sexual size dimorphism is not well understood, compared to that in adults. The present study investigates the ontogeny of sexual size dimorphism in Saunders's gulls (Larus saundersi) in captivity. We artificially incubated fresh eggs collected in Incheon, South Korea, and measured body size, locomotor activity, and foraging skill in post-hatching chicks in captivity. Our results indicated that the sexual differences in size and locomotor activity occurred with the post-hatching development. Also, larger males exhibited greater foraging skills for food acquisition than smaller females at 200 days of age. Future studies should assess how the adaptive significance of the sexual size dimorphism in juveniles is linked with sexual divergence in survival rates, intrasexual contests, or parental effort in sexes.

We assessed dispersal and vicariant events in four species of Japanese moles in the genera Mogera and Euroscaptor to better understand the factors shaping intra- and interspecific differentiation in Japanese moles. We used the combined viewpoints of molecular phylogeny and historical geology using nucleotide sequences of mitochondrial (cytochrome b; Cytb) and nuclear (A2ab, Bmp4, Tcf25, vWf) genes. The divergence times estimated from the molecular data were verified with available geological data on the chronology of fluctuations in sea level in the Korea Strait, assuming sequential migration and speciation events. This produced possible migration times of 5.6, 3.5, 2.4, and 1.3 million years ago for four species of Japanese moles, Euroscaptor mizura, Mogera tokudae, M. imaizumii, and M. wogura, respectively. For the western Japanese mole M. wogura, Cytb sequences revealed four major phylogroups with strong geographic affinities in southwestern Central Honshu (I), western Honshu/Shikoku (II), Kyushu/westernmost Honshu (III), and Korea/Russian Primorye (IV). The nuclear gene sequences supported the distinctiveness of phylogroups I and IV, indicating long, independent evolutionary histories. In contrast, phylogroups II and III were merged into a single geographic group based on the nuclear gene data. Intraspecific divergences in M. imaizumii and M. tokudae were rather apparent in Cytb but not in nuclear gene sequences. The results suggest that repeated dispersal events have occurred between the Asian continent and the Japanese Islands, and intensive vicariant events associated with abiotic and biotic factors have created higher levels of species and genetic diversities in moles occurring on the Japanese Islands.

Geographic ranges of parthenogenetic animals often differ from those of their sexual conspecifics, providing valuable information about evolutionary histories. We studied the geographic distributions of sexual and parthenogenetic forms of the weevil Scepticus insularis (Roelofs) (Coleoptera, Curculionidae) on Honshu, Shikoku, and Kyushu Islands, Japan. We collected Scepticus insularis at 18 sites in the northern half of Honshu; sequenced 633 bp of the mitochondrial ND2 (NADH dehydrogenase subunit 2) gene for 102 Honshu beetles; and constructed a ML gene tree based on these sequences and sequences previously reported for 104 conspecific beetles from Hokkaido of known reproductive mode. The ND2 haplotypes fell into two distinct clades (I and II) separated by 9.5% nucleotide sequence divergence. The haplotypes from all parthenogenetic females on Hokkaido and those from all-female populations on Honshu invariably belonged to clade I, whereas sexual females on Hokkaido and some of the females in male/female populations on Honshu belonged to clade II. Haplotypes of males were in clade II, except for two Hokkaido males having a clade-I haplotype. Excluding these two enigmatic males, we treated individuals with clade-I haplotypes as parthenogenetic, and those with clade-II haplotypes as sexual. Parthenogenetic beetles were widespread throughout Hokkaido and central and northern Honshu, whereas sexuals were found only in three separate areas on Hokkaido and one on Honshu, often sympatrically with parthenogens. The distribution of S. insularis is congruent with the pattern termed geographic parthenogenesis.

Insects in many species vary greatly in the expression of secondary sexual traits, resulting in sexual dimorphism, which has been proposed to be a consequence of differences in sexual selection. In this study, we analyze the occurrence of sexual shape dimorphism and the correlation between geometric body size in males and females and sex ratio in the genus Ceroglossus (Coleoptera: Carabidae) using geometric morphometrics and randomization analysis. Our results show a positive relationship between the centroid size of males and females and sex ratio. However, we also observed a trend to maintain a “similar” body shape between sexes and species due to characteristics associated with sexual selection of the group in which the scarcity of outstanding morphological characters does not restrict reproduction capacity. The trend in sexual proportions was found to be near 1:1 for the eight species. We propose that the observed differences between sexes associated with the sex ratio are due to an energy cost caused by sexual selection; nevertheless, since proportions are similar for all the species, it is really not relevant to generate more conspicuous structures for a stronger male-male competence.

The four major Chinese carps are commercially important fish species with high production in China. However, their recruitment decreased sharply in the Yangtze River since the late 20^th century. In the present study, to reveal the relationships between spawning activities of the four species and environmental factors, drifting eggs were collected at Yidu City, in the middle reaches of the Yangtze River, from May to July each year between 2005 and 2010. Classification and regression trees (CART) analysis was applied to identify the key factors associated with spawning activities of the four carp species. Twelve predictor variables (hydrological and meteorological variables) and one response variable (egg presence or number of egg) were included in the CART. Our CART analysis showed that water temperature and the diurnal increase of water level were the two most significant factors for the spawning activities. When water temperature was between 18°C and 24°C, especially in association with the diurnal increase of water level greater than 0.55 m·d^-1, spawning activities was always favored. Unlike the hydrological factors, meteorological factors seemed to have no influence on initiating the spawning activities. The density of drifting eggs of the four species was mainly influenced by the diurnal variation of water level, the diurnal variation of water discharge, water temperature, humidity, and air pressure. We then related our results to the ecological regulation of the Three Gorge Reservoir in the middle reaches of the Yangtze River. We suggested that, when water temperature was between 18–24°C from May to July, to ensure the successful spawning of the four carp species, the ecological regulation should be managed to create flood peaks and make the diurnal increase of water level greater than 0.55 m·d^-1.

The circannual pupation rhythm of Anthrenus verbasci is entrained to an environmental cycle by changes in photoperiod. Exposure of larvae reared under short-day conditions to long days induced a clear phase delay of the circannual rhythm. There was no notable difference in the initial phase or period of the circannual rhythm among four geographically distinct populations of A. verbasci in Japan: Takanabe (32.1°N), Osaka (34.7°N), Sendai (38.3°N), and Sapporo (43.1°N) populations. The range of photoperiodic changes effective for phase delay in the circannual pupation rhythm was compared among the four populations. Although larvae did not show a typical threshold response, but responded quantitatively to the photophase duration in intermediate conditions, the critical daylengths were calculated as those under which the pupation was delayed by 50%: 12.8 h in the Takanabe population, 13.2 h in the Osaka population, and 13.6 h in the Sendai and Sapporo populations. Thus, the critical daylength for entrainment of the circannual rhythm in A. verbasci was correlated to the habitat latitude, but the differences among the populations were much smaller than those reported in photoperiodism for induction of diapause in various insects. Consequently, the difference in the pupation time among the four geographic populations was very small under the natural photoperiod in Osaka at 20°C, and absent under the natural photoperiod and temperature in Osaka. These results suggest that A. verbasci survives and successfully produces the next generation in different geographic regions without changing the parameters of the circannual rhythm.

Three forms of gonadotropin-releasing hormone (GnRH) are found in vertebrates; these differ in amino acid sequence, localization, distribution, and embryological origin. We used northern blot analysis, and in situ hybridization to detect GnRH transcripts in various tissues in the large ascidian Halocynthia roretzi. We cloned a cDNA encoding two novel GnRHs, termed tGnRH-10 and tGnRH-11, from H. roretzi, with deduced amino acid sequences of QHWSYGFSPG and QHWSYGFLPG, respectively. Both GnRHs are highly similar to those of teleosts and tetrapods. For example, the tGnRH-10 sequence is 90% identical to seabream GnRH1, and tGnRH-11 is 90% identical to salmon GnRH3. The primary structure of the deduced preprotein is similar to that of chordate GnRHs and consists of a signal peptide, two decapeptides, up- and downstream processing sequences (containing lysine and arginine), and a GnRH-associated peptide. The transcripts of the H. roretzi GnRH gene were expressed in all tissues examined. Comparison of the signal peptide of the lamprey GnRH-II precursor with those of three forms from representative vertebrates revealed homology to GnRH2 precursors. These novel ascidian GnRHs offer a new perspective on the origin of vertebrate GnRH subtypes. We hypothesize that gnathostome GnRH2 was derived only from lamprey GnRH-II and that ancestral gnathostome GnRH, which produces neurons that originate in peripheral organs, gave rise to vertebrate GnRH1 and GnRH3 through whole-genome duplication.

We describe the morphology and innervation of the midgut and hindgut of the giant land snail Megalobulimus abbreviatus for the first time. The midgut (stomach and intestine) and hindgut (rectum and anus) are innervated by the subesophageal ganglia, through the gastrointestinal branch (originated from the visceral nerve) and the rectum-anal nerve, respectively. Backfilling through these nerves revealed neuronal bodies, mainly in the right parietal and visceral ganglia. The enteric plexuses of the midgut and hindgut are formed by extensive axonal networks and several neuronal somata arranged in clusters or as isolated cells. The gastrointestinal branch and the rectum-anal nerve directly innervate the enteric plexuses of the intestine and the hindgut, respectively. However, the outer wall of the stomach has a stomatogastric nervous system, which consists of four ganglia: stomatogastric, gastric, cardic, and pyloric. Fibers of the gastrointestinal branch project to these ganglia. Anterograde tracing from stomatogastric system ganglia revealed that the enteric plexus of the stomach is innervated only by these peripheral ganglia. Anterograde tracing of the gastrointestinal branch did not result in labeling in the enteric plexus of the stomach. Therefore, the midgut and hindgut of M. abbreviatus is controlled by an intrinsic innervation, constituted by the submucous and myenteric plexuses, which are innervated directly by neurons from the subesophageal ganglia or indirectly via the stomatogastric nervous system (for the stomach).

Xenos is a strepsipteran genus whose members are parasitic to eusocial wasps, including the hornet genus Vespa. We undertook an extensive sampling of strepsipterans in Xenos from hornets collected in East Asia and performed molecular phylogenetic analyses based on mitochondrial cytochrome c oxidase subunit I gene sequences (652 bp) to investigate the cryptic diversity among 21 individuals of strepsipterans. The analyses, accompanied by morphological examination, revealed that these strepsipterans represent two distinct species, X. moutoni du Buysson, 1903 and X. oxyodontes sp. nov. The two species differed in their host-utilization pattern: the latter was almost specific to Vespa analis and V. simillima, whereas the former was associated with other species in Vespa.