Multiple spawning of amphidromous type ayu <Emphasis Type="Italic">Plecoglossus altivelis</Emphasis> in a large river,Mogami River System

Female ayu Plecoglossus altivelis were sampled from the Mogami River System, Yamagata Prefecture to investigate the occurrence of multiple spawning that has previously been reported in small rivers. Spawning frequencies of the females were estimated by histological observations of the ovary mainly from the degree of degeneration of postovulatory follicles. In the middle and late part of the spawning period, most females showed characteristics of having spawned two or three times, indicating that multiple spawning is a common phenomenon also in a large river. Mean relative batch fecundity (RBF) and condition factor (without gonads) decreased with the number of spawning batches. Size dependency in the occurrence of the multiple spawning and the RBF values was not observed. These results are similar to those reported in previous studies of ayu from a small river near the Mogami River System, and indicate that the occurrence and properties of multiple spawning are almost the same regardless of the river scale. However, it is possible that the ratios of multiply spawned females may differ due to variations in survival rates during the spawning season.     

Residence pattern of the ayu <Emphasis Type="Italic">Plecoglossus altivelis</Emphasis><Emphasis Type="Italic">altivelis</Emphasis> larvae and juveniles occurring in the surf zone of a sandy beach,Niigata Prefecture,northern Sea of Japan

This study examined the residence pattern of ayu larvae and juveniles occurring in the surf zone of an extensive sandy beach, in the north part of the Sea of Japan. In this region, ayu larvae mainly hatched in October and November, generally appeared in the surf zone in hatching order, and were continuously present until the following January. During a short period after mid- to late January when the water temperature fell below 10°C, ayu larvae and juveniles concurrently migrated from the surf zone regardless of their size and age, resulting in a longer term residence for earlier-hatched ayu. This was in contrast to studies in other waters, which found that early-hatched ayu have a short-term residence and late-hatched a long-term residence. Furthermore, faster growth that enables ayu to reach the maximum residence size (around 40 mm BL) in the surf zone during the period when the water temperature was still over 10°C might lead to earlier migration and a shorter residence term. This process might be applicable only to the early-hatched group (defined as individuals hatched in late September and October) and depends on the size of each individual, thus leading to a gradual migration of larger individuals as they attain maximum residence size.     

Molecular cloning and expression analysis of <Emphasis Type="Italic">Megalobrama amblycephala</Emphasis> transferrin gene and effects of exposure to iron and infection with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

Transferrin (Tf) plays an important function in iron homeostasis and metabolism of organisms. In this study, we identified and characterized the Tf gene in Megalobrama amblycephala and evaluated its expression in basal conditions as well as after iron overload and experimental infection with Aeromonas hydrophila. Furthermore, we studied the iron binding properties of recombinant Tf. The full-length M. amblycephala Tf complementary DNA (cDNA) (GenBank accession no.: KX698308) of 2245 bp was cloned and contained a 1953 bp open reading frame (ORF) encoding 650 amino acid residues and flanked by a 68 bp 5′ and a 204 bp 3′ untranslated regions (UTR). Predicted conservative structure illustrated that M. amblycephala Tf consisted of two conservative Tf domains. Amino acid sequence alignment revealed that M. amblycephala Tf had high similarity with that of cyprinids deposited in Genbank, and phylogenetic analysis showed that M. amblycephala Tf clustered with Ctenopharyngodon idella and Hypophthalmichthys molitrix. Tissue expression pattern analyses demonstrated that the liver was the main Tf mRNA expressing organ, being significantly higher than other tissues (p < 0.05). In the liver, Tf mRNA expression in fish artificially injected with the pathogenic bacteria A. hydrophila was significantly upregulated, reaching a peak at 12 h post injection (hpi) and then decreasing afterward. The expression in FeCl₃-injected fish showed a similar tendency, but reached a peak at 8 hpi. Meanwhile, fish serum iron significantly decreased following A. hydrophila injection, but increased to peak at 4 hpi and then decreased in FeCl₃-injected fish. The recombinant M. amblycephala Tf showed iron binding capacity using CAS analysis. These results are helpful to understand the structure and regulation of expression of Tf, as well as the specific function of Tf for both immune responses and iron homeostasis.     

Identification and expression analysis of ghrelin gene in common carp <Emphasis Type="Italic">Cyprinus carpio</Emphasis>

ABSTRACT:   A ghrelin gene has been cloned and sequenced in common carp Cyprinus carpio . Ghrelin cDNA is composed of 461 bp [with a 36-bp 5'-untranslated region (UTR) and a 113-bp 3'-UTR], which translates into a protein of 103 amino acid residues. Carp ghrelin (preproghrelin) contained a predicted signal peptide of 26 amino acid residues, the ghrelin domain ( Gly ²⁷– Val ⁴⁵) and C-terminal peptide ( Gly ⁴⁶– Phe ¹⁰³). Homology analysis of the ghrelin domain of carp with that of other known ghrelin in vertebrates showed good similarity to teleost ghrelin (50–81.8%). Hydropathy analysis based on the deduced amino acid sequence of ghrelin domains in teleosts showed a similar profile. Carp ghrelin clustered with ghrelin of goldfish Carassius auratus and other teleosts, away from mammalian, reptilian, avian, amphibian and chondrichthian ghrelin, by phylogenetic analysis. Genomic organization of carp ghrelin gene was composed of four exons and three introns, which was the same as that of other teleosts and human ghrelin genes. The carp ghrelin gene was expressed in unstimulated tissues such as foregut, hindgut, spleen and brain. In spleen cells, expression of the ghrelin gene increased upon stimulation with lipopolysaccharide (LPS), phytohemagglutinin (PHA) or imiquimod. The identification of carp ghrelin gene and the analysis of the modulation of its expression in immune-activated conditions will allow a more complete analysis of the roles of ghrelin in teleosts.     

Identification of Wap65, a human homologue of hemopexin as a copper-inducible gene in swordtail fish, <Emphasis Type="Italic">Xiphophorus helleri</Emphasis>

Copper is one of the major heavy metal pollutants found in the aquatic environment. Therefore, it is important for determining the genes that play a key role in copper metabolism in aquatic organisms. This study, thus, aimed to identify a new copper-inducible gene in swordtail fish, Xiphophorus helleri. Using ACP-based RT-PCR coupled with RLM-RACE, we cloned Wap65, a mammalian homologue of hemopexin gene. The gene exhibits high identity at amino acid levels with the Wap65 gene of other fish species (42–68%) and mammalian hemopexin gene (35–37%). In addition, ten cysteine and two histidine residues are conserved in the swordtail fish Wap65 gene. These cysteine residues are vital for structural integrity, and histidine residues provide high binding affinity towards heme. As revealed by RT-PCR, the gene was upregulated in swordtail fish that were exposed to copper in a dose- and time-dependent manner. Therefore, the identification of Wap65, a mammalian homologue of hemopexin, as a new copper-inducible gene will provide greater insight into the role of this gene in copper metabolism.