Maternal transcripts in good and poor quality eggs from Japanese eel, Anguilla japonica-their identification by large-scale quantitative analysis.

Our understanding of maternal control of development in vertebrates remains incomplete. In this study, we investigated levels of maternal transcripts in good and poor quality eggs from artificially matured Japanese eel, using RNA-Seq and quantitative polymerase chain reaction (qPCR), to identify candidate maternal transcripts related to development. De novo assembly or mapping of reads to the eel draft genome yielded 619,029 contigs and 85,906 transcripts, respectively; normalized read counts to these assemblies were calculated using reads (RPKM) or fragments (FPKM) per kilobase of transcript per million mapped reads. In silico screening identified 1,594 contigs and 150 transcripts with lower RPKM or FPKM in poor than in good quality eggs, 245 contigs, and 85 transcripts of which could be annotated by BLASTx, respectively. From selected contigs or transcripts, six genes (dnajb4, gnpat, card14, pdp1, fcgbp, ttn) had significantly lower messenger RNA levels in poor than in good quality eggs by qPCR. Multiple regression analysis showed that five genes (gnpat, b4galnt1, acsl6, rtkn, trim24) significantly correlated with hatchability. Taken together, 10 genes were identified as candidate maternal transcripts, regulating development in Japanese eel. Our results contribute to understanding the molecular basis for maternal control of development in vertebrates.

Anti-Oxidant and Fucoxanthin Contents of Brown Alga Ishimozuku (Sphaerotrichia divaricata) from the West Coast of Aomori, Japan.

Fucoxanthin is a specific carotenoid in brown seaweeds with remarkable biological properties. Ishimozuku (Sphaerotrichia divaricata), an edible brown alga from northern Japan, has morphology that is almost identical to that of Okinawa-mozuku (Cladosiphon okamuranus) harvested off Okinawa, Japan. However, because of Ishimozuku's lower availability compared to Okinawa-mozuku, the contents of its nutrient compounds remain unclear. The present study analyzed fucoxanthin and anti-oxidant compound contents of Ishimozuku harvested off the northern coast of Japan from 2014 to 2016. First, 80% ethanol extract solutions were prepared from Ishimozuku harvested from several west coast areas of Aomori, Japan. Then, polyphenol content was analyzed using the Folin⁻Ciocalteu method. Then anti-oxidative effects were analyzed by their 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and hydrogen peroxide scavenging activity. Furthermore, fucoxanthin contents were measured using high performance liquid chromatography (HPLC) analysis. Fucoxanthin contents of Ishimozuku were 105.6⁻1148.5 µg/g dry weight. Total polyphenol contents of Ishimozuku were of 0.296⁻0.958 mg/g dry weight: higher than Okinawa-mozuku (0.082 ± 0.011 mg/g dry weight). The anti-oxidation effects of Ishimozuku accompanied the polyphenol content. These results suggest that Ishimozuku contains various anti-oxidant components and has high potential to provide the promotion of human health.

Histological Evidence of Multiple Spawning in Wild Female Japanese Eel Anguilla japonica.

The post ovulatory follicle (POF) is an important and reliable tissue structure used to investigate the spawning history in teleost fish. Fresh POFs shortly after spawning are comprised of cellular (follicular cells) and acellular (basement membrane and fibrils such as elastic fibers) components. The cellular components are quickly disintegrated by means of apoptosis, while the acellular components persist for a longer period. Since cellular components are well visualized by conventional hematoxylin-eosin (HE) staining but acellular components are not stained well, old POFs that have lost cellular components are difficult to identify. In this study, periodic acid-Schiff and Victoria blue staining, which can distinctly visualize acellular POF components, were applied to the ovarian tissues of Japanese eel (Anguilla japonica) (n = 9) captured from June to August of 2008, 2009, and 2013 at the southern West Mariana Ridge, a spawning area for Japanese eels. Only new POFs were observed in seven females caught in June, and these females had ovaries with early-to mid-vitellogenic stage oocytes. Both fresh and old POFs were observed in a female caught in July, and only mid-vitellogenic stage oocytes were observed. Only old POFs and no vitellogenic stage oocyte were observed in a female caught in August. A progressive decrease in muscle lipid content, gonad somatic index, and condition factors was observed from June to August. Thus, the female Japanese eel can spawn at least twice or three times at most during spawning season, depending on energy reserve.

Oral Administration of Edible Seaweed Undaria Pinnatifida (Wakame) Modifies Glucose and Lipid Metabolism in Rats: A DNA Microarray Analysis.

SCOPE: Wakame is an edible seaweed that is a common constituent in the Japanese diet. Previous studies showed that wakame consumption is associated with the prevention of metabolic syndrome, but the molecular mechanisms underlying the protective effects are poorly understood. METHODS AND RESULTS: To determine if the expression of hepatic genes is affected by ingestion of the brown seaweed Undaria pinnatifida (wakame), rats were fed a diet containing 0, 0.1, or 1.0 g per 100 g dried wakame powder for 28 days. Administration of 1% wakame significantly decreased serum total cholesterol levels. Hepatic gene expression was investigated using DNA microarray analysis, and the results showed that wakame suppresses the lipogenic pathway by downregulating SREBF-1. Moreover, bile acid biosynthesis and gluconeogenesis were promoted by upregulation of the PPAR signaling pathway, which leads to a reduction in the accumulation of cholesterol and promotion of ß-oxidation. CONCLUSIONS: These results suggest that wakame ingestion affects glucose and lipid metabolism by altering the expression of SREBF-1 and PPAR signal-related genes.

Gonadal soma-derived factor (gsdf), a TGF-beta superfamily gene, induces testis differentiation in the teleost fish Oreochromis niloticus.

The Nile tilapia, Oreochromis niloticus, is a gonochoristic teleost fish with an XX/XY genetic system and is an excellent model for gonadal sex differentiation. In the present study, we screened novel genes that were expressed predominantly in either XY or XX undifferentiated gonads during the critical period for differentiation of gonads into ovaries or testes using microarray screening. We focused on one of the isolated 12 candidate genes, #9475, which was an ortholog of gsdf (gonadal soma-derived factor), a member of the transforming growth factor-beta superfamily. #9475/gsdf showed sexual dimorphism in expression in XY gonads before any other testis differentiation-related genes identified in this species thus far. We also overexpressed the #9475/gsdf gene in XX tilapia, and XX tilapia bearing the #9475/gsdf gene showed normal testis development, which suggests that #9475/gsdf plays an important role in male determination and/or differentiation in tilapia.