Two estrogen receptors expressed in the teleost fish, Sparus aurata: cDNA cloning, characterization and tissue distribution.
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Estrogen is an essential hormone for many reproductive and non-reproductive functions. The function of estrogen in the reproductive cycle of seabream (Sparus aurata), a protandrous hermaphrodite teleost fish, is complex but it is understood to be involved in sex inversion, a process that occurs in some individuals during the second reproductive season. Estrogen action is mediated by two estrogen receptor (ER) subtypes designated alpha and beta. As a step to understanding the mechanisms of estrogen action during natural and induced sex reversal in seabream, we have isolated two cDNAs encoding distinct forms of ER homologous to mammalian ERalpha and ERbeta. The seabream ERalpha clone (sbERalpha1), which was truncated in the A/B domain, corresponded to a variant differing in five amino acids from another recently cloned sbERalpha. The ERbeta clone (sbERbeta) encoded a protein 559 amino acids long and showed only 40% identity to sbERalpha. Northern blot analysis of liver and ovary mRNA indicated the presence of several transcripts of the two receptor subtypes. PCR analysis showed that the two receptors differed in their expression pattern. sbERalpha had a more restricted distribution, occurring mainly in testis, liver and heart, and sbERbeta was present in most tissues, being more abundant in ovary, testis, liver, intestine and kidney. The presence in seabream of two ERs with several ER transcripts and their pattern of distribution are consistent with the widespread effects of estrogen in different tissues. (+info)
alpha-MSH acetylation in the pituitary gland of the sea bream (Sparus aurata L.) in response to different backgrounds, confinement and air exposure.
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MSH is a pituitary hormone derived by post-translational processing from POMC and involved in stress and background adaptation. N-terminal acetylation of MSH to monoacetyl alpha-MSH or diacetyl alpha-MSH increases the bioactivity of the peptide. The aim of this study was to characterize alpha-MSH acetylation in the sea bream (Sparus aurata L.) pituitary gland in response to the stressors air exposure and confinement, as well as in fish adapted for 15 days to a white, gray or black background. Pituitary homogenates were purified by reversed-phase HPLC (RP-HPLC). The alpha-MSH content of fractions was measured by RIA. Immunoreactive RP-HPLC fractions were further analyzed by electrospray mass spectrometry and the peptide sequence determined as SYSMEHFRWGKPV-NH2. In the pituitary gland of sea bream, des-, mono- and diacetyl alpha-MSH were identified. Then plasma alpha-MSH levels were measured in sea bream adapted to different backgrounds. Surprisingly, we found the highest plasma alpha-MSH levels in white-adapted as compared with black-adapted sea bream with intermediate values for gray-adapted fish. This observation is in contrast with results that have been obtained in eel, trout or terrestrial vertebrates. Next, des-, mono- and diacetyl alpha-MSH forms were measured in homogenates of the pituitary gland and in plasma of sea bream exposed to air, to confinement, or to different backgrounds. Monoacetyl alpha-MSH was the predominant form in all control and experimental groups. The lowest content of monoacetyl alpha-MSH relative to des- and diacetyl alpha-MSH was found in white-adapted fish. Levels of des- and diacetyl alpha-MSH forms were similar under all conditions. We observed that monoacetyl alpha-MSH is the most abundant isoform in the pituitary gland after background adaptation, confinement and air exposure, in sea bream. These data indicate that the physiologically most potent isoform of alpha-MSH may vary from species to species. (+info)
Glucokinase gene expression is nutritionally regulated in liver of gilthead sea bream (Sparus aurata).
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Glucose intolerance in carnivorous fish has been attributed to the lack of hepatic glucokinase (GK) activity. Transcription/translation assay and transient transfection of COS-7 cells with a cDNA encoding Sparus aurata liver GK showed the functionality of the enzyme in vitro. The endogenous fish hepatic GK had lower affinity for glucose than the rat enzyme. The GK activity values in fed fish were similar to those reported for starved and diabetic rats. In this study, we also addressed the nutritional regulation of GK gene expression in fish liver. Starvation and energy restriction decreased S. aurata hepatic GK mRNA and activity levels, as previously reported in rats. In contrast, the fish enzyme expression exhibited a delayed onset during the daily feeding rhythm. These findings demonstrate for the first time the presence and the nutritional modulation of a functional GK activity in fish liver and contribute to explain the low ability of carnivorous fish to metabolize carbohydrates. (+info)
Molecular cloning and hormonal control in the ovary of connexin 31.5 mRNA and correlation with the appearance of oocyte maturational competence in red seabream.
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Gap junctions are aggregates of intercellular channels, composed of the protein connexin (Cx), between adjacent cells. This study examined whether, in the ovary of the red seabream Pagrus major, the connexin gene essential for the production of RNA and protein during the acquisition of oocyte maturational competence is active. Mixed primers for this reaction were designed on the basis of the high sequence homology of selected regions of known connexin genes. Polymerase-chain-reaction-amplified cDNA fragments generated by 3' and 5' rapid amplication of cDNA ends were combined to generate full-length cDNA sequences. The resulting 2400 base pair cDNA had an open reading frame encoding a polypeptide containing 275 amino acid residues (31493 Da; Cx31.5). Hydropathicity analysis of the predicted amino acid sequence indicated that red seabream Cx31.5 has four major hydrophobic regions and four major hydrophilic regions indicative of a topology similar to that of known connexins. Typical connexin consensus sequences were also observed in the first and second extracellular loops. During the acquisition of oocyte maturational competence, red seabream Cx31.5 mRNA transcription levels increased after treatment with gonadotropin-II. It is therefore proposed that expression of Cx31.5 contributes to the acquisition of oocyte maturational competence in this species. (+info)
The relationship between the position of the retinal area centralis and feeding behaviour in juvenile black bream Acanthopagrus butcheri (Sparidae: Teleostei).
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The topography of the neurons in the retinal ganglion cell layer of juvenile black bream Acanthopagrus butcheri changes during development. The region of high cell density the area centralis (AC), relocates from a temporal (central) to a dorsal (peripheral) position within the dorso-temporal retinal quadrant. To ascertain whether the differences in the position of the AC during development are related to feeding behaviour, we monitored fishes that were given a choice of food. A range of feeding behaviour patterns was recorded in individual fishes. The smallest fishes (8-15 mm standard length (SL)) took live food from the water column. Following weaning onto pellets, fishes exhibited a preference for taking food from either the substrate or the surface (but not both). When greater than 20 mm SL, a number of individuals then divided their time between surface and substrate feeding before all fishes became exclusive benthic feeders at a stage between 50 and 80 mm SL. Three individual fishes, for which behaviour patterns were categorized, were killed and the topography of the retinal ganglion cell layer analysed. A range of positions for the AC was found with the smallest fish (12 mm SL) possessing a region of high cell density in the temporal retina. In a larger fish (70 mm SL), feeding from both the substrate and the surface, the AC was found in an intermediate dorso-temporal position. The AC of a fish (51 mm SL) preferentially taking food from the substrate was located in a dorsal position. (+info)
Empirical Bayes procedure for estimating genetic distance between populations and effective population size.
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We developed an empirical Bayes procedure to estimate genetic distances between populations using allele frequencies. This procedure makes it possible to describe the skewness of the genetic distance while taking full account of the uncertainty of the sample allele frequencies. Dirichlet priors of the allele frequencies are specified, and the posterior distributions of the various composite parameters are obtained by Monte Carlo simulation. To avoid overdependence on subjective priors, we adopt a hierarchical model and estimate hyperparameters by maximizing the joint marginal-likelihood function. Taking advantage of the empirical Bayesian procedure, we extend the method to estimate the effective population size using temporal changes in allele frequencies. The method is applied to data sets on red sea bream, herring, northern pike, and ayu broodstock. It is shown that overdispersion overestimates the genetic distance and underestimates the effective population size, if it is not taken into account during the analysis. The joint marginal-likelihood function also estimates the rate of gene flow into island populations. (+info)
Histopathology of cultured sea bream Sparus aurata infected with sanguinicolid trematodes.
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The present study is the first report of a sanguinicolid infection affecting sea bream Sparus aurata cultured in net cages in the NE of Spain. The disease was associated with trickling mortalities during the cold season (1999 and 2000). Examination of gill wet mounts of the affected population revealed that sanguinicolid infection was present in 82.6 and 100% of the fish sampled in 1999 and 2000, respectively. Adult flukes, which were located in the kidney, were tentatively identified as members of the family Sanguinicolidae, subfamily Cardicolinae. Eggs and miracidia were found in the gill vascular structures. The inflammatory response triggered by the parasites was moderate and the lesions caused by either eggs and miracidia in the gills or adult flukes in the kidney were not extremely severe, possibly because of the moderate intensity of the parasitosis. Histological observations of sanguinicolid infected sea bream presented here are compared with those reported in other fish species. The role played on sea bream morbility and mortality by other factors (occurrence of a simultaneous moderate monogenean infection, immunological impairement related to low water temperatures) is discussed. (+info)
Predictive modeling of the shelf life of fish under nonisothermal conditions.
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The behavior of the natural microflora of Mediterannean gilt-head seabream (Sparus aurata) was monitored during aerobic storage at different isothermal conditions from 0 to 15 degrees C. The growth data of pseudomonads, established as the specific spoilage organisms of aerobically stored gilt-head seabream, combined with data from previously published experiments, were used to model the effect of temperature on pseudomonad growth using a Belehradek type model. The nominal minimum temperature parameters of the Belehradek model (T(min)) for the maximum specific growth rate (micro(max)) and the lag phase (t(Lag)) were determined to be -11.8 and -12.8 degrees C, respectively. The applicability of the model in predicting pseudomonad growth on fish at fluctuating temperatures was evaluated by comparing predictions with observed growth in experiments under dynamic conditions. Temperature scenarios designed in the laboratory and simulation of real temperature profiles observed in the fish chill chain were used. Bias and accuracy factors were used as comparison indices and ranged from 0.91 to 1.17 and from 1.11 to 1.17, respectively. The average percent difference between shelf life predicted based on pseudomonad growth and shelf life experimentally determined by sensory analysis for all temperature profiles tested was 5.8%, indicating that the model is able to predict accurately fish quality in real-world conditions. (+info)