Embryonic expression of UCP2 in rainbow trout (Oncorhynchus mykiss)

Uncoupling proteins are mitochondrial anions transporters that dissociate respiration from ATP synthesis through proton leaks. Uncoupling protein 2 reportedly plays a role in several physiological processes such as energy partitioning, nutrition, and fatty acid metabolism. The mRNA expression of rainbow trout UCP2 genes (UCP2A and UCP2B) was monitored during embryogenesis and early larval development. Both genes were recruited early and displayed similar steadily decreasing patterns from fertilization until hatching. The expression of UCP2A and UCP2B appeared significantly differentiated after hatching and during the yolk sac absorption, with UCP2A displaying higher expression. We suggest that UCP2 expression profiles in the rainbow trout embryo could be associated with the utilization of lipids as a source of energy during development.     

The effects of elevated winter temperature and sub-lethal pollutants (low pH, elevated ammonia) on protein turnover in the gill and liver of rainbow trout (Oncorhynchus mykiss)

Appetite, growth, and protein turnover (synthesis, growth and degradation) of liver and gills were measured in juvenile rainbow trout (Oncorhynchus mykiss) fed to satiation, and exposed for 90 days to elevated winter temperatures (+2 °C above ambient) and either low pH (5.2) in softwater or 70 M total ammonia (TAmm) in hardwater. All fish increased in weight during the experiments, but those exposed to +2°C grew significantly more than those at ambient temperature due to a stimulation of appetite. During the relatively constant temperature of the first 75 days, +2 °C caused a significant increase in the rates of protein synthesis and degradation in the liver of hardwater-acclimated fish, as a result of an increase in RNA translational efficiency (KRNA). The elevated temperature also induced an increase in gill protein synthesis in softwater-acclimated fish but in this case the underlying mechanism was an increase in Cs, the capacity for protein synthesis (RNA:protein) rather than in KRNA. The addition of 70 M TAmm had no effect on protein turnover in either liver or gills of hardwater-acclimated fish. Low pH inhibited protein growth in the liver of softwater-acclimated fish at day 90 under both temperature regimes. This inhibition was effected via a decrease in protein synthesis at control temperature but via an increase in protein degradation when the fish were exposed to both low pH and +2 °C. From these results we conclude that a simulated global warming scenario has potentially beneficial rather than detrimental effects on protein turnover and growth of freshwater fish during winter.     

Protein kinase C in the spleen of the turbot (Scophthalmus maximus L.)

PKC activity was detected in spleen extracts from the turbot, Scophthalmus maximus, a teleost flatfish that is farmed commercially in several countries, in assays with the substrate EGF- R651–658 as phosphate acceptor. The activity was purified about 700-fold by a three-step chromatographic procedure (DEAE-cellulose, phenyl-Sepharose and threonine-Sepharose). Maximal activity was obtained in the presence of the typical PKC cofactors Ca2+ (0.1 mM) PtdS (20 g ml–1) and either DAG (2 g ml–1) or PMA (2 g ml–1). Activity was dose-dependently inhibited by H7 and by the PKC-specific inhibitors PKC19–36 and N-myristoylated PKC19–31. The rate of phosphorylation was highest with the PKC-specific substrate MARCKS161–175. In immunoblotting, MC5 (a mouse monoclonal antibody raised against bovine PKC) recognized bands of 80 and 100 kDa. Immunoblotting with antibodies raised against mouse PKC isozymes (, , , , , , and ) indicated the presence of all these isozymes in turbot spleen.     

Binding characteristics of the Zn-binding membrane protein from common carp

ABSTRACT:    This study incorporated the 43 kDa Zn-binding membrane protein isolated from common carp into liposome. The specificity and strength of the binding of ⁶⁵Zn to the 43 kDa protein-liposomes, and the binding of the ⁶⁵Zn-labeled 43 kDa protein-liposomes to laminin were studied. It was found that ⁶⁵Zn was bound to the external side of the 43 kDa protein-liposomes. Specific binding of ⁶⁵Zn to the protein-liposomes was detected. The binding parameter of Zn to the protein was found to be: maximum binding site (N_max), 76.7 pmole/µg protein (approx. 3 mole of Zn²⁺/mole); and equilibrium dissociation constant (Kd), 0.19 µM. Of the cations introduced (Ca²⁺, Cd²⁺,Co²⁺, Cr²⁺, Cu²⁺, Fe²⁺, Hg²⁺, Mg²⁺, Mn²⁺, Ni²⁺, Pb²⁺), only Co²⁺ competed significantly with Zn. The protein-liposomes were also found to bind specifically to laminin with a N_max of 1.1 pmole/µg laminin, and Kd of 4.79 µM. No significant protein-liposome binding occurred to other extracellular matrix proteins (fibronectin, fibrinogen or vitronectin). Furthermore, the binding was specifically inhibited by the Arg-Gly-Asp (RGD) peptide or GRGDSPG, while two other analogs (GRGESPG and GRADSPG) were without effect.     

Polka dot grouper Cromileptes altivelis fingerlings require high protein and moderate lipid diets for optimal growth and nutrient retention

An 8‐week comparative slaughter experiment was carried out to determine the effect of dietary protein and lipid on growth, apparent digestibility (AD) and nutrient retention of polka dot grouper Cromileptes altivelis. Fingerlings were fed diets that varied in crude protein (CP) at 55 g kg^?1 increments between 410 and 630 g kg^?1 dry matter (DM) and at either a moderate (150 g kg^?1 DM) or high (240 g kg^?1DM) lipid concentration. Each diet was fed to satiety twice daily to four replicate tanks (110 L) of fish. One replicate block of tanks comprised 150 fish of mean (±SD) initial weight of 9.6 ± 0.29 g, which were distributed equally to 10 tanks. The other three replicate blocks of tanks comprised 300 fish of 12.6 ± 0.45 g, which were distributed equally to 30 tanks. Tanks were provided with filtered and heated (29 ± 0.5 °C) seawater in a flow‐through system within a laboratory where photoperiod was maintained at 12 : 12 h light–dark cycle. Voluntary food intake was not significantly affected by either the CP or lipid concentration of the diet (mean ± SD of 1.93 ± 0.146 g week^?1) but there was a trend for intake to be higher on the moderate compared with the high lipid diets (mean ± SEM of 1.97 versus 1.89 ± 0.033 gweek^?1, respectively). Daily growth coefficient (DGC) and food conversion ratio (FCR) improved linearly (P < 0.01) with increasing dietary CP (from 0.94 to 1.35% day^?1 for DGC and 1.58 to 1.00 g DM g^?1 wet gain for FCR) and these responses were almost coincident for each of the lipid series. The AD of CP increased linearly with increasing dietary CP (from 46.8 to 74.1%) and was independent of dietary lipid. Apparent digestibility of energy increased curvilinearly with increasing dietary CP, with the quadratic component being more prominent for the high‐lipid series. Increasing the amount of lipid in the diet markedly increased the lipid content of the fish from an initial composition (mean ± SD) of 173 ± 7.3 g kg^?1 to a final composition (mean ± SEM) of either 217 or 250 ± 5.9 g kg^?1 for moderate and high‐lipid series, respectively. Total body lipid content tended to increase linearly with increasing dietary CP for the high‐lipid series but with an opposite effect for the moderate‐lipid series. The retention of digestible nitrogen decreased linearly with increasing dietary CP but at a steeper rate for the moderate, compared with the high, lipid series (from 62.7 to 35.7%, slope ?0.115 for moderate‐lipid and 54.6 to 41.9%, slope ?0.050 for high‐lipid). A quadratic function of dietary CP concentration best explained the retention of digestible energy with the curvilinearity being more marked for the high, compared with the moderate, lipid diet series. While there was some indication that ingested lipid spared dietary protein, the results showed a far greater propensity of polka dot grouper fingerlings to use protein as the prime dietary energy source. Diets for juvenile polka dot grouper should contain not less than 440 g digestible protein kg^?1 DM and at least 150 g lipid kg^?1 DM.     

Minimising aerobic respiratory demands could form the basis to sub-lethal copper tolerance by rainbow trout gill epithelial cells in vitro

Mechanisms of Cu tolerance were investigated in respiratory epithelial cell cultures, from rainbow trout gills, by studying O₂ consumption and protein synthesis rates, intracellular Na concentration and TER. The lowest concentration found to reduce O₂ consumption was 25 M Cu. This did not affect either protein synthesis rate or intracellular Na concentration and was interpreted in terms of copper tolerance; i.e., how these two energetically demanding processes are maintained despite a reduction in aerobic ATP supply. The relationship between protein synthesis rate and synthesis cost is exponential and the cost of protein synthesis in gill cells was found to be minimal (i.e., this cell occupies a position on the asymptotic section of the protein synthesis rate/synthesis cost model) and unaffected by 25 M Cu. Thus protein synthesis rates could be maintained since any reduction would represent an insignificant energy saving. Intracellular Na concentrations and O₂ consumption rates were linearly correlated suggesting reducing intracellular maintenance costs would have a greater significance in terms of overall energetic conservation. Intracellular Na maintenance costs, calculated from O₂ consumption rates and intracellular Na concentrations, were found to decline after exposure to 25 M Cu. Since TER was unaffected this implied the reduced costs arose from membrane `channel arrest'. Thus the Na/K ATPase energy demands, associated with maintaining intracellular Na concentration, could be reduced by decoupling metabolic demand and membrane function. Therefore this study may demonstrate how the flexibility of cellular energetics enables gill epithelial cells to tolerate sub-lethal Cu.     

玛曲渔场几种裂腹鱼类脑的形态结构与真食性的相互关系

本文对极边扁咽齿鱼,黄河裸裂尻鱼、花斑裸鲤和厚唇重唇鱼进行了食性分析和脑的形态结构观察,并对脑结构和食性的关系作了初步探讨。认为这四种鱼类属于底层味觉食性鱼类,其中极边扁咽齿鱼和黄河裸裂尻鱼主要靠口腔、腭部味蕾觅食泥土和藻类,迷叶发达;厚唇重唇鱼主要靠触须寻找甲壳类为主的水底昆虫,面叶和小脑发达;花斑裸鲤的脑结构特征与其杂食性相适应。     

Global-scale genetic structure of a cosmopolitan cold-water coral species

• 1. When considering widely distributed marine organisms with low dispersal capabilities, there is often an implication that the distribution of cosmopolitan species is an artefact of taxonomy, constrained by the absence of characters for delimiting either sibling or cryptic species. Few studies have assessed the relationship among populations across the global range of the species' distribution, and the presence of oceanographic barriers that might influence gene flow among populations are underestimated.
• 2. In this study, evolutionary and ecological drivers of connectivity patterns have been inferred among populations of the cold-water coral Desmophyllum dianthus, a common and widespread solitary scleractinian species, whose reproduction strategy and larval dispersal are still poorly unknown.
• 3. The genetic structure of D. dianthus was explored using 30 microsatellites in 347 specimens from 13 localities distributed in the Mediterranean Sea and Atlantic and Pacific Oceans.
• 4. Results clearly reveal genetically differentiated populations in the Northern and Southern Hemispheres (F_ST = 0.16, F_SC = 0.01, F_CT = 0.15, P-values highly significant), and Chilean and New Zealand populations with independent genetic profiles.
• 5. Marine connectivity patterns at different spatial scales are discussed to characterize larval dispersal and gene flow through the Northern and Southern Hemispheres.

     

Genetic identification of native populations of fluvial white-spotted charr <Emphasis Type="Italic">Salvelinus leucomaenis</Emphasis> in the upper Tone River drainage

ABSTRACT:   Stocking of exogenous, hatchery-reared white-spotted charr Salvelinus leucomaenis has been conducted throughout much of their range in Honshu Island, Japan, to increase angling opportunities. Although the native charr populations are thought to have declined because of hybridization with introduced fish, their distribution and genetic status have been uncertain. Fine population structures of charr in the upper Tone River drainage were examined using mitochondrial DNA and microsatellite analyses so as to clarify the presence of native populations. One common mtDNA haplotype was detected in all populations in the Ohashi River and Watarase River, and four and one tributary populations were monomorphic for such haplotypes, respectively. However, several haplotypes, considered to have originated from stocked hatchery fish, were observed in the stocked and the remaining populations. Judging from the genetic integrity over a fine geographic scale, the former were considered as indicative of native populations and the latter as admixtures with hatchery fish. Comparisons of genetic diversity, deviations from the Hardy–Weinberg equilibrium, principal component analysis, and relatedness estimations based on microsatellite DNA can also provide evidence for distinguishing native populations from those influenced by hatchery fish.     

Characterization of a cDNA encoding a homolog of actin-related protein 4 from a marine red alga Porphyra yezoensis

ABSTRACT:   A cDNA ( PyARP4 ) containing an open reading frame for a protein of 573 amino acids was identified in the marine red alga Porphyra yezoensis . The conceptual PyARP4 protein exhibits significant similarity to actin-related protein (ARP) 4 in the terrestrial plant Arabidopsis . Comparison of the deduced amino acid sequence showed moderate sequence identity (30%) to a conventional actin in P. yezoensis , as seen in comparisons between ARP and conventional actins of other organisms. A putative bipartite nuclear localization signal and an actin motif were found within the PyARP4 amino acid sequence. In a phylogenetic analysis, the PyARP4 was found to cluster with the ARP4 of other organisms. The expression level of PyARP4 did not change significantly among four developmental stages of life cycle and was lower than that of a conventional actin. This cDNA therefore may serve as a useful internal standard in gene expression analyses of differentially expressed genes in P. yezoensis .