Cloning of kuruma prawn Marsupenaeus japonicus crustin-like peptide cDNA and analysis of its expression

ABSTRACT:   Antimicrobial peptides serve as an important component of the innate immune system of all species by functioning to provide a rapid first line defense against infection. Arthropod antimicrobial peptides have been well described in insects, whereas only a few molecules have been identified in crustaceans. Five variants (types 1–5) of Marsupenaeus japonicus crustin-like peptide cDNA that were obtained from a hemocyte cDNA library and polymerase chain reaction (PCR) amplification are reported here. Marsupenaeus japonicus crustin-like peptide type 1, the predominant type, has a cDNA consisting of 679 nucleotides and an open reading frame consisting of 573 base pairs coding for 191 amino acid residues. Other types contain varying glycine-rich repeats at the N-terminal amino acid sequences. The deduced amino acid sequences of these variants are highly similar to those of Litopenaeus setiferus (80% identity), Litopenaeus vannamei (80% identity) and Carcinus maenas crustins (44% identity). Expression of Marsupenaeus japonicus crustin-like peptide mRNA was detected in hemocytes, but not in the heart, hepatopancreas, gill, fore-gut, mid-gut, muscle, subcuticular epithelium or ovary. The expression level of crustin-like peptide mRNA increased significantly 1, 3 and 7 days post-peptidoglycan feeding as determined by quantitative real-time PCR. These results suggest that crustin-like peptide could have an important role in shrimp defense mechanisms.     

Effects of temperature and food availability on growth rate during late larval stage of Japanese anchovy (Engraulis japonicus) in the Kuroshio–Oyashio transition region

During periods when the population size of Japanese anchovy Engraulis japonicus is large, the abundance of 1‐yr olds has been considered to be dependent on the growth and survival processes in the late larval and early juvenile stages in the Kuroshio–Oyashio transition region off northern Japan. Recent growth rates for 10 days before capture of larval and early juvenile E. japonicus were estimated and examined in relation to the surface water temperature and the available copepod density in 1997, 1998 and 1999. Late larval and early juvenile E. japonicus were distributed in the waters with temperature from 15 to 19°C and available prey density from 10 to 1000 mg dry weight (DW) m^?2 in the transition region. The late larval growth rates were found to be regulated more strongly by water temperature than by copepod density in the waters <16°C, and more strongly by copepod density than water temperature in the waters <100 mg DW m^?2 in the Kuroshio–Oyashio transition region. The recent growth rates decreased from the western waters to the eastern waters in the survey area 140–170°E in 1998, correlating with decreases of food availability to 50–100 mg DW m^?2. While in 1999, the recent growth rates were faster in the waters east of 150°E, resulting from eastward expansion of warm water ranges and high available prey density 100–400 mg DW m^?2. The key environmental factors regulating late larval growth rate of E. japonicus in the transition region seem to be spatially different between years.     

Isolation of microsatellite markers by in silico screening implicated for genetic linkage mapping in Japanese pufferfish Takifugu rubripes

ABSTRACT:   The search for dinucleotide repeat microsatellites within scaffolds 1–25 of genome database JGI Fugu v3.0 for the pufferfish Takifugu rubripes revealed that 80% of microsatellite loci consisted of five to 13-fold repeats with locus-specific differences in density. Eleven out of 15 microsatellite loci isolated from the database with which genotyping using wild pufferfish was successfully performed showed polymorphism; that is, the means of the number of alleles and expected and observed heterozygosities at these 11 loci were 21.8, 0.915 and 0.829, respectively. It was confirmed that eight out of the 11 polymorphic loci were inherited through the Mendelian law and one pair of microsatellite loci derived from the same scaffold was linked. These results demonstrated that these loci are useful for constructing a linkage map in the pufferfish as DNA markers.     

Prolonged exposure to hyaluronidase decreases the fertilization and development rates of fresh and cryopreserved mouse oocytes

Hyaluronidase is generally used to remove cumulus cells from mouse oocytes before oocyte cryopreservation, intracytoplasmic sperm injection or DNA injection. In general, use of cumulus-free mouse oocytes decreases in vitro fertilizing ability compared with cumulus-surrounded oocytes. The effect of hyaluronidase exposure on the quality of mouse oocytes is not fully understood. Here, we investigated the effect of hyaluronidase exposure time on the fertilization rate of fresh and vitrified mouse oocytes and their subsequent developmental ability in vitro. We found that the fertilization rate decreased with hyaluronidase treatments. This reduction in the fertilization rate following treatment with hyaluronidase was fully reversed by removal of the zona pellucida. In addition, oocytes treated with hyaluronidase for 5 min or longer had a reduced capacity to develop to the morula and blastocyst stage. The survival, fertilization, and developmental rates of vitrified-warmed oocytes were also reduced by longer exposure to hyaluronidase. In conclusion, these results suggest that prolonged exposure to hyaluronidase decreases the quality of mouse oocytes and shorter hyaluronidase treatment times may help achieve a stable and high fertilization rate in fresh and cryopreserved oocytes.     

Comparative study of morphological changes in hardwoods treated with the ionic liquid, 1-ethyl-3-methylimidazolium chloride

Three hardwoods of varying vessel arrangement were treated with the ionic liquid, 1-ethyl-3-methylimidazolium chloride ([C2mim][Cl]), which can dissolve cellulose, to investigate its influence on wood tissue morphology. Characterization was carried out by light and scanning electron microscopy. The wood fibers of all species swelled significantly during [C2mim][Cl] treatment. The swelling behavior varied according to wood species and differed from other cell types such as ray parenchyma cells. Morphological changes of the pits also varied between wood species. Treatment with [C2mim][Cl] affects wood tissues differently depending on wood species and cell type. These differences are not due to the vessel arrangement and its presence, but possibly from the chemical component and the microfibril angle of various wood tissues.     

Occurrence of Spontaneous Tumors in the Central Nervous System (CNS) of F344 and SD Rats

In order to accurately assess the carcinogenicity of chemicals with regard to rare tumors such as rat CNS tumors, sufficient information about spontaneous tumors are very important. This paper presents the data on the type, incidence and detected age of CNS tumors in F344/DuCrlCrlj (a total of 1363 males and 1363 females) and Crl:CD(SD) rats (a total of 1650 males and 1705 females) collected from in-house background data-collection studies and control groups of carcinogenicity studies at our laboratory, together with those previously reported in F344 and SD rats. The present data on F344/DuCrlCrlj rats (F344 rats) and Crl:CD(SD) rats (SD rats) clarified the following. (1) The incidences of all CNS tumors observed in F344 rats were less than 1%. (2) The incidences of malignant astrocytoma and granular cell tumor were higher in male SD rats than in female SD rats. (3) The incidences of astrocytoma and granular cell tumor were higher in SD rats than in F344 rats. (4) Among astrocytoma, oligodendroglioma and granular cell tumor, oligodendroglioma was detected at the youngest age, followed by astrocytoma, and ultimately, granular cell tumor developed in both strains. The incidences observed in our study were almost consistent with those previously reported in F344 and SD rats.     

Successive use of different habitats during the early life stages of Pacific herring <Emphasis Type="Italic">Clupea pallasii</Emphasis> in Akkeshi waters on the east coast of Hokkaido

Distributions of eggs, larvae and juveniles of Pacific herring Clupea pallasii were surveyed in Lake Akkeshi and the connecting Akkeshi Bay on the Pacific coast of eastern Hokkaido. Eggs were found attached to seagrasses and seaweeds in the densely vegetated eastern inner lake. Larvae (7.1–34.9 mm total length) were distributed in the less densely vegetated inner lake during April and June. Juveniles (35.0–89.6 mm) were collected in the central and western parts of the lake. The sites containing juveniles were less densely vegetated with water temperatures lower than 20 °C from June to August. When the water temperature of the entire lake rose to 20 °C in late summer, juveniles appeared to move from the lake to the bottom layer of Akkeshi Bay, which has lower temperatures. Thus, in Akkeshi waters, C. pallasii successively used different habitats during its egg, larval and juvenile stages. A comparison of the current limited distribution of eggs and larvae with the distribution over the entire lake and bay areas in the 1950s and 1960s (periods of large catch size) indicates that the spawning grounds and larval habitats of C. pallasii have contracted to the lake area due to low spawning stock biomass in recent years.