Increased liver protein and mRNA expression of natural killer cell-enhancing factor B (NKEF-B) in ayu (Plecoglossus altivelis) after Aeromonas hydrophila infection.

Natural killer cell-enhancing factor (NKEF) may mediate cellular responses to proinflammatory molecules. The liver proteins of Aeromonas hydrophila-infected ayu (Plecoglossus altivelis) and healthy control fish were analyzed by 2DE. A protein, which increased significantly in diseased fish, was identified as NKEF-B by MALDI-TOF-MS. A full-length cDNA clone of this proteinwas subsequently isolated. It contains 1092 bp with an open reading frame of 591 bp, coding for 197 amino acids with MW 21.9 kDa and pI 6.38, values similar to those determined by 2DE. Ayu NKEF-B had highest similarity (93.1% amino acid identity) to those of carp and zebrafish. Phylogenetic analysis showed that ayu NKEF-B falls into the fish NKEF-B cluster and is most closely related to that of carp and zebrafish. It was determined that ayu NKEFB mRNA expression was significantly increased in many tissues at the early stage of bacterial infection. In conclusion, the increased NKEF-B mRNA and protein expression in ayu were closely associated with A. hydrophila infection.

The effects of environmental salinity on trunk kidney proteome of juvenile ayu (Plecoglossus altivelis).

As the life cycle of ayu spans river, brackish and seawater environments, it would be a suitable fish model for studying the responses to salinity changes in aquatic animals. We investigated the effect of salinity on trunk kidney proteome in ayu (Plecoglossus altivelis) using two-dimensional gel electrophoresis and mass spectrometry. The proteins involved in the process of energy metabolism, biosynthesis, DNA methylation and cell differentiation were mainly affected, and 10 significantly changed proteins were identified. Our result showed that isocitrate dehydrogenase (ICD), pyruvate dehydrogenase (E1), O-glycosyl hydrolase, mitochondrial precursor of ATP synthase subunit beta, mitochondrial ferrtin (MtF), retinol binding protein (RBP) were down-regulated, whereas aldehyde dehydrogenase, cytokeratin 1, S-adenosylhomocysteine hydrolase, Cys-Met metabolism PLP-dependent enzyme were up-regulated when ayu transferred from freshwater to brackish water. Partial coding sequences of E1, ICD, MtF and RBP genes were determined, and the effects of salinity on their mRNA expression in ayu trunk kidney were tested by real-time PCR subsequently. Their possible direct or indirect roles in the adaptation of ayu to salinity are discussed.