Zoological Science

Myosin-binding protein-C (MyBP-C), also known as C-protein, is a major myosin-binding protein characteristic of striated muscle, and plays a critical role in myofibril organization, especially in registration of thick filaments in the sarcomeres during myofibrillogenesis. We previously demonstrated that cardiac-type MyBP-C is involved early in the process of myofibrillogenesis in both cardiac and skeletal muscle during chicken muscle development. Two variants (type I and type II) have been detected in chicken cardiac MyBP-C; they differ only in the presence or absence of a sequence of 15 amino acid residues (termed P-seq) that includes a phosphorylation site for cyclic AMP-dependent kinase in the cardiac MyBP-C motif (Yasuda et al, 1995). Therefore, types I and II are regarded as phosphorylatable and non-phosphorylatable isoforms, respectively. In this study, an antibody specific for P-seq was prepared. With this and other monoclonal antibodies to cardiac MyBP-C (C-315), expression and localization of the two MyBP-C isoforms in developing chicken cardiac and skeletal muscle were examined by immunocytochemistry and immunoblotting. The results showed that type I is predominantly expressed in the heart and is localized in myofibrils of both atrial and ventricular muscles through development. In contrast, type II is mainly expressed in embryonic skeletal muscle, although type I is faintly expressed in cultured skeletal muscle. These observations were confirmed by RT-PCR.

Cryptic species are common in gobioid fishes, as revealed by recent molecular studies. This study collected molecular and morphological data to delimit species boundaries and to reveal the historical biogeography of Odontamblyopus lacepedii sensu lato by sampling 87 specimens from 16 locations in the northwestern Pacific. Phylogenetic trees based on mitochondrial NADH dehydrogenase subunit 5 sequences identified three major clades. Clear morphological separations among these clades were detected by examining morphological characters used in taxonomic diagnosis. Therefore, these clades were delineated as three separate species: O. lacepedii sensu stricto (in the East China Sea, Yellow Sea, Gulf of Bohai, and Ariake Bay); O. sp. (in the southern East China Sea and southern Yellow Sea); and O. rebecca (in the Gulf of Tonkin, northern South China Sea, and southern East China Sea). Estimated divergence times were 0.61 ± 0.15 Ma between O. lacepedii and O. sp., and 2.31±0.36 Ma between O. rebecca and O. lacepedii+O. sp. These findings indicate that isolation of marginal seas and habitat fragmentation during major falls in sea level in the late Pliocene and Pleistocene may have been responsible for genetic breaks among Odontamblyopus species in the northwestern Pacific.

Mitochondrial DNA sequence polymorphisms and patterns of genetic diversity represent the genealogy and relative impacts of historical, geographic, and demographic events on populations. In this study, historical patterns of population dynamics and differentiation in hawksbill (Eretmochelys imbricata) and green turtles (Chelonia mydas) in the Pacific were estimated from feeding populations in the Yaeyama Islands, Japan. Phylogenetic relationships of the haplotypes indicated that hawksbill and green turtles in the Pacific probably underwent very similar patterns and processes of population dynamics over the last million years, with population subdivision during the early Pleistocene and population expansion after the last glacial maximum. These significant contemporary historical events were suggested to have been caused by climatic and sea-level fluctuations. On the other hand, comparing our results to long-term population dynamics in the Atlantic, population subdivisions during the early Pleistocene were specific to Pacific hawksbill and green turtles. Therefore, regional differences in historical population dynamics are suggested. Despite limited sampling locations, these results are the first step in estimating the historical trends in Pacific sea turtles by using phylogenetics and population genetics.

Activating transcription factor / cyclic-AMP response element-binding protein (ATF/CREB) has been implicated as a key regulator in the transcriptional control of many genes. In this study, we isolated and characterized a full-length cDNA that encodes a CRE-binding protein 2 (CREB2) called ATF4 in Xenopus embryos. Like other CREB 2 transcription factors, the 342-amino acid ATF4 protein contains a carboxyl terminal leucine-zipper motif, an adjacent basic domain, and an amino terminal leucine-zipper motif. Unlike other CREB2 (ATF4) proteins, the ATF4 isolated from the gonads of Xenopus embryos contains a consensus phosphorylation site for protein kinase A (PKA). In a gel shift analysis, ATF4 bound to a CLS sequence in the promoter of Xenopus aromatase.

Reproductive activities of salmonids are synchronized by changes in photoperiod, which control the endocrine system via the brain-pituitary-gonadal axis. Gonadotropin-releasing hormone (GnRH) in the brain regulates synthesis and release of the pituitary gonadotropins (GTHs; FSH and LH). FSH and LH in turn stimulate the production of sex steroids for oocyte growth and maturation-Inducing steroid hormones for oocyte maturation and ovulation, respectively, in female salmonids. To clarify effects of long-term photoperiod manipulations on the reproductive activity of salmonids from early recrudescence to ovulation, we Investigated the gene expression profiles of GnRH, GTHs, and vitellogenin (VTG), and plasma sex steroids in female rainbow trout (Oncorhynchus mykiss). In addition, the percentages of eyed embryos and hatched alevins were examined together with the number of ovulated eggs to evaluate the effects of photoperiod regimes on egg quality. During late summer, the mRNA levels of GnRHs, GTHα, and LHβ, and the plasma level of a maturational steroid (17α,20β-dihydroxy-4-pregnen-3-one; 17,20β-P) were significantly elevated by a gradually shortened photoperiod under constant temperature, in accordance with accelerated sexual maturation. The percentages of eyed embryos and hatched alevins from fish ovulated in August were comparable to those of control fish observed in December. These results clearly indicate that syntheses of GnRHs, LH, VTG, and 17,20β-P are effectively accelerated by a programmed long-short photoperiod regime in early recrudescent female rainbow trout, without a marked deterioration in egg quality.

We conducted an electrophoretic survey to examine geographic genetic variation in samples from 17 localities of the Japanese clawed salamander, Onychodactylus japonicus. This species was divided into six genetic groups (N-Tohoku, S-Tohoku, Tsukuba, SW-Honshu, Kinki, and Shikoku) that were largely concordant with clades or subclades recognized in our previous mtDNA study. Although the relationships among these six groups were not clarified, genetic distances between them were not small (mean Nel's D=0. 146–0.471). Among these groups, the geographically isolated Tsukuba group was genetically distinct, possibly as a result of population isolation. In a locality of western Honshu, two groups, SW-Honshu and Shikoku, were found to occur sympatrically. Although several presumable hybrid individuals were found, hybridization between these two groups seemed to occur very rarely. These results indicate that the Shikoku group is specifically distinct from the SW-Honshu group, whose range includes the type locality of O. japonicus.

Leptin, an adipocyte-derived hormone, plays important roles in metabolism and reproduction. In this article, we report the cloning, expression, and identification of the giant panda leptin (gLeptin) gene and its variants. The gLeptin cDNA was 504 bp long, encoding a precursor peptide of 167 amino acids including 21 residues of signal peptide. A short variant of gLeptin was 501 bp long, encoding a 166-aa peptide and also including a 21-aa signal peptide. Giant panda leptin was 99.4%, 94.6%, and 92.8% identical to that of black bear, dog, and cat, respectively, but was only 81.4% and 80.8% identical to that of human and rat. The cloned gLeptin gene was expressed in Escherichia coli, with expression confirmed by Western blotting and MALDI-TOF-TOF MS PMF. After purification, renaturation, and condensation, the gLeptin protein was injected into Kunming mice. The recombinant gLeptin significantly inhibited food intake by 41.8% and reduced body weight by 5.1% in the mice.

The aim of this study was to investigate consecutive stages of gonadal development of the yellow-bellied toad (Bombina variegata) with particular emphasis on the origin of somatic and germ cell lineages as well as the timing of gonial cell migration. Changes in gonadal basal lamina distribution helped to explain the exceptional mode of gonadal differentiation in this species. Atypical and rapid differentiation of the male gonad in B. variegata is the result of the ability of gonial cells to migrate into the center of the gonad relatively early. Thus, the testis medulla contains germ cells from the onset of gonadal differentiation into cortex and medulla, whereas in other anurans a sterile medulla is characteristic of both future testes and ovaries; germ cells translocate into the medulla during the subsequent stage of testis development. This atypical testiculogenesis is probably the result of an acceleration of the sex determination period, indicating a contribution of sex determination heterochrony to the course of gonadogenesis. The results also suggest that medullar cells are derived from proliferating coelomic epithelial cells. Moreover, Sertoli cells constitute an integral part of the germinal epithelium in B. variegata, as in other vertebrates. Spermatids do not contact Sertoli cells just before spermiation and do not form bundles.

Four species of the coelotine spider genus Himalcoelotes Wang, 2002 endemic to the Himalayas were studied: H. gyirongensis (Hu and Li, 1987); H. tortuous sp. nov.; H. xizangensis (Hu, 1992); and H. zhamensis sp. nov. The male of H. xizangensis is here described for the first time. A key to all 13 Himalcoelotes species is given. Twelve Himalcoelotes species are divided into two species groups. The species-group placement of H. aequoreus Wang, 2002 is uncertain.

We describe a new species of torrent-dwelling ranid frog of the genus Meristogenys from the Crocker Range, western Sabah, northern Borneo. The new species, Meristogenys maryatiae, differs from congeners by the combination of: small body, males 31–37 mm and females 65–66 mm in snout-vent length; head narrower than long; eyes moderate, diameter subequal to snout; iris unicolored; legs long; ventral surface of tibia without heavy pigmentation; rear of thigh blotched dark brown and cream; toes fully webbed; outer metatarsal tubercle present; larval dental formula 7(4–7)/6(1).