Browsing by Author "Mancera, J. M."
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- 16.P15. Branchial osmoregulatory response to salinity challenge in the Lusitanian toadfishPublication . Guerreiro, Pedro M; Laiz-Carrión, R.; Haond, Christophe; Modesto, Teresa; Fuentes, Juan; Mancera, J. M.; Canario, Adelino V. M.Halobatrachus didactylus, a marine teleost found in coastal lagoons and river estuaries is often exposed to important salinity changes. Despite its aglomerular kidney, it is able to survive in hypo-osmotic environments, likely via compensatory actions from gills and intestine. We aimed at evaluating the response of the branchial tissue of H. didactylus to salinity changes.
- AVT is involved in the regulation of ion transport in the intestine of the sea bream (Sparus aurata)Publication . Martos-Sitcha, J. A.; Gregorio, Silvia; Carvalho, Edison Samir Mascarelhas; Canario, Adelino V. M.; Power, Deborah; Mancera, J. M.; Martínez-Rodriguez, G.; Fuentes, J.The intestine of marine fish plays a crucial role in ion homeostasis by selective processing of ingested fluid. Although arginine vasotocin (AVT) is suggested to play a role in ion regulation in fish, its action in the intestine has not been demonstrated. Thus, the present study investigated in vitro the putative role of AVT in intestinal ion transport in the sea bream (Sparus aurata). A cDNA encoding part of an AVT receptor was isolated and phylogenetic analysis revealed it clustered with the V1a2-type receptor clade. V1a2 transcripts were expressed throughout the gastrointestinal tract, from esophagus to rectum, and were most abundant in the rectum regardless of long-term exposure to external salinities of 12, 35 or 55 p.p.t. Basolateral addition of AVT (10 6 M) to the anterior intestine and rectum of sea bream adapted to 12, 35 or 55 p.p.t. mounted in Ussing chambers produced rapid salinity and region dependent responses in short circuit current (Isc), always in the absorptive direction. In addition, AVT stimulation of absorptive Isc conformed to a dose–response curve, with significant effects achieved at 10 8 M, which corresponds to physiological values of plasma AVT for this species. The effect of AVT on intestinal Isc was insensitive to the CFTR selective inhibitor NPPB (200 lM) applied apically, but was completely abolished in the presence of apical bumetanide (200 lM). We propose a role for AVT in the regulation of ion absorption in the intestine of the sea bream mediated by an absorptive bumetanide-sensitive mechanism, likely NKCC2.
- Branchial osmoregulatory response to salinity in the gilthead sea bream,Sparus auratusPublication . Laiz-Carrión, R.; Guerreiro, P. M.; Fuentes, J.; Canario, Adelino V. M.; Martín Del Río, María P.; Mancera, J. M.The branchial osmoregulatory response of gilthead sea bream (Sparus auratus L.) to short-term (2–192 hr) and long-term (2 weeks) exposure to different environmental salinities (5%, 15%, 25%, 38% and 60%) was investigated. A ‘‘U-shaped’’ relationship was observed between environmental salinity and gill Naþ,Kþ-ATPase activity in both long- and short-term exposure to altered salinity, with the increase in activity occurring between 24 and 96 hr after the onset of exposure. Plasma osmolality and plasma ions (sodium, chloride, calcium and potassium) showed a tendency to increase in parallel with salinity. These variables only differed significantly (Po0.05) in fish adapted to 60% salinity with respect to fish adapted to full-strength sea-water (SW). Plasma glucose remained unchanged whereas plasma lactate was elevated at 5% and 60%. Muscle water content (MWC) was significantly lower in fish adapted to 60%. Chloride cells (CC) were only present on the surface of the gill filaments and absent from the secondary lamellae. CC distribution was not altered by external salinity. However, the number and size of CC were significantly increased at salinity extremes (5% and 60%), whereas fish exposed to intermediate salinities (15% and 25%) had fewer and smaller cells. Furthermore, the CC of fish exposed to diluted SW became rounder whereas they were more elongated in fish in full-strength and hypersaline SW. This is consistent with previous reports indicating the existence of two CC types in euryhaline fish. At likely environmental salinities, gilthead sea bream show minor changes in plasma variables and the effective regulation of gill Naþ,Kþ-ATPase. However, at very low salinities both haemodilution and up-regulation of gill Naþ,Kþ-ATPase predict a poor adaptation most likely related to deficiency or absence of specific components of the CC important for ion uptake.
- Changes in plasma amino acid levels in a euryhaline fish exposed to different environmental salinitiesPublication . Aragão, C.; Costas, B.; Vargas-Chacoff, L.; Ruiz-Jarabo, I.; Dinis, Maria Teresa; Mancera, J. M.; Conceição, L. E. C.Previous studies have shown that Senegalese sole is partially euryhaline in the juvenile phase, being able to adapt to a wide range of salinities in a short-time period, due to changes at the osmoregulatory and metabolic level. This study aimed to assess the effects of acclimation of sole to a wide range of salinities, with a special emphasis on the role of plasma amino acids during this process. Sole juveniles were acclimated for 2 weeks to different salinities: 5, 15, 25, 38, and 55 g L−1. Plasma levels of cortisol, glucose, osmolality, and free amino acids were assessed at the end. Changes in plasma levels of cortisol, glucose, and amino acids indicate that fish reared at 5 and 55 g L−1 were facing extra energy costs. Amino acids seem to play an important role during salinity acclimation, either as energy sources or as important osmolytes for cell volume regulation.
- Characterization of the peripheral thyroid system of gilthead seabream acclimated to different ambient salinitiesPublication . Ruiz-Jarabo, I.; Klaren, P. H. M.; Louro, Bruno; Martos-Sitcha, J. A.; Pinto, P. I.; Vargas-Chacoff, L.; Flik, G.; Martinez-Rodriguez, G.; Power, Deborah; Mancera, J. M.; Arjona, F. J.Thyroid hormones are involved in many developmental and physiological processes, including osmoregulation. The regulation of the thyroid system by environmental salinity in the euryhaline gilthead seabream (Sparus aurata) is still poorly characterized. To this end seabreams were exposed to four different environmental salinities (5, 15, 40 and 55 ppt) for 14 days, and plasma free thyroid hormones (fT3, ff4), outer ring deiodination and Na+/K+ -ATPase activities in gills and kidney, as well as other osmoregulatory and metabolic parameters were measured. Low salinity conditions (5 ppt) elicited a significant increase in fT3 (29%) and ff4 (184%) plasma concentrations compared to control animals (acclimated to 40 ppt, natural salinity conditions in the Bay of Cadiz, Spain), while the amount of pituitary thyroid stimulating hormone subunit 13 (tshb) transcript abundance remained unchanged. In addition, plasma fT4 levels were positively correlated to renal and branchial deiodinase type 2 (dio2) mRNA expression. Gill and kidney T4-outer ring deiodination activities correlated positively with dio2 mRNA expression and the highest values were observed in fish acclimated to low salinities (5 and 15 ppt). The high salinity (55 ppt) exposure caused a significant increase in tshb expression (65%), but deiodinase gene expression (diol and dio2) and activity did not change and were similar to controls (40 ppt). In conclusion, acclimation to different salinities led to changes in the peripheral regulation of thyroid hormone metabolism in seabream. Therefore, thyroid hormones are involved in the regulation of ion transport and osmoregulatory physiology in this species. The conclusions derived from this study may also allow aquaculturists to modulate thyroid metabolism in seabream by adjusting culture salinity. (C) 2016 Elsevier Inc. All rights reserved.
- Dietary aflatoxin B1 (AFB1) reduces growth performance, impacting growth axis, metabolism, and tissue integrity in juvenile gilthead sea bream (Sparus aurata)Publication . Barany, A.; Guilloto, M.; Cosano, J.; de Boevre, M.; Oliva, M.; de Saeger, S.; Fuentes, J.; Martinez-Rodriguez, G.; Mancera, J. M.Mycotoxins are an increasing threat to all the related commodities from agriculture. Its occurrence is expected to increase due to climate change. Here, we examined the impacts of dietary toxicity of aflatoxin B1 (AFB1) in gilthead sea bream (Sparus aurata) at levels of 1 or 2 mg AFB1 kg(-1)- fish feed. Inclusion of AFB1 in the diet resulted in 80% inhibition of the total weight gain during the 85-day trial. Carbohydrate and lipid energetic metabolites, both in plasma and liver, were depleted. Moreover, the histopathological analysis revealed several tissue anomalies in the liver, kidney, and spleen. Furthermore, the relative expression of gene transcripts for growth regulation was affected by AFB1. Adenohypophyseal gh and hepatic igf1 were inversely correlated due to AFB1 effects. Relative expression levels of gene transcripts as stress indicators were increased at AFB1 highest doses, such as hypothalamic trh, crh, and crhbp, as well as star in head kidney. Interestingly circulating levels of cortisol were unaffected. Overall, our results showed that aquafeeds with AFB1 impaired growth, alter metabolism, tissue integrity, and transcriptomic responses. However, we did find AFB1 residue neither in the liver nor muscle.
- Different environmental temperatures affect amino acid metabolism in the eurytherm teleost Senegalese sole (Solea senegalensis Kaup, 1858) as indicated by changes in plasma metabolitesPublication . Costas, B.; Aragão, C.; Ruiz-Jarabo, I.; Vargas-Chacoff, L.; Arjona, F. J.; Mancera, J. M.; Dinis, Maria Teresa; Conceição, L. E. C.Senegalese sole (Solea senegalensis) is a eurytherm teleost that under natural conditions can be exposed to annual water temperature fluctuations between 12 and 26C. This study assessed the effects of temperature on sole metabolic status, in particular in what concerns plasma free amino acid changes during thermal acclimation.
- Divergence of duplicate POMC genes in gilthead sea bream Sparus auratusPublication . Cardoso, João; Laiz-Carrión, R.; Louro, Bruno; Silva, Nádia; Canario, Adelino V. M.; Mancera, J. M.; Power, DeborahProopiomelanocorticotrophin (POMC) in vertebrates is produced in the pituitary gland and undergoes post-translational processing to give rise to a range of biologically active peptides. Teleosts possess 2–3 different POMC transcripts which have been proposed to have originated from a whole or partial genome duplication. In the present study 2 transcripts of gilthead sea bream POMC (sbPOMC-a1 and a2) were cloned and characterised. sbPOMC-a1 is expressed principally in the melanotroph cells of the pars intermedia (PI) and sbPOMC-a2 is expressed in the corticotroph cells of the rostral pars distalis and probably also in the PI. The 2 sbPOMC transcripts have a differential tissue distribution in extra-pituitary sites. An appraisal of POMC evolution indicates sbPOMCs belong to one of the two main clades that exist in teleosts and that overall a non conservative process of gene loss occurred in this infraclass.
- Effect of amino acid supplementation and stress on expression of molecular markers in meagre (Argyrosomus regius)Publication . Herrera, M.; Matias, A. C.; Soares, F.; Ribeiro, L.; Moreira, Márcio; Salamanca, N.; Jerez-Cepa, I.; Mancera, J. M.; Astola, A.The objectives of this work were: 1) develop of molecular stress biomarkers obtaining sequence data of different transcripts, 2) study the molecular stress response through the expression quantification of key gene involved in it, and 3) assess the effects of dietary amino acid additives on stress response in meagre meagres (Argyrososmus regius). Fish batches were fed two experimental diets with tryptophan (Trp) or aspartate (Asp) added for seven days. Before sampling fish were submitted to confinement/netting stress during 1 h, except control fish. Therefore fish were sampled before and after stress (1 h and 6 h post-stress). The sampling consisted of blood and tissues (brain, hypophysis and liver). Several gene expressions related to the stress response were measured in those tissues, and the cloning of corticotropin-releasing hormone (crh), corticotropin-releasing hormone binding protein (crh-bp), and thyrotropin-releasing hormone (trh) has been reported in meagre for the first time. In fact, fish fed an additional Asp diet did not present any sl, prl and gh expression changes, as for the control group. Contrarily, the Trp diet altered the prl and gh expressions after stress. For crh and crh-bp expressions, no significant differences were detected within the Asp diet hence that amino acid improved the stress response. However, Asp feeding, but not Trp, enhanced pomc-a expression after stress. Hsp70 expression varied for every treatment, including the control feeding, indicating a late response at 6 h post-stress sampling, where both Asp and Trp treatments increased these expressions significantly. Concluding, the response of molecular stress markers to amino acid enriched diets was diverse. The stressor did not change significantly the relative expression of most analyzed genes for control feeding groups, though the Asp supplemented diet was more effective for attenuating molecular markers than the Trp one.
- Expression of pituitary prolactin, growth hormone and somatolactin is modified in response to different stressors (salinity, crowding and food-deprivation) in gilthead sea bream Sparus auratusPublication . Laiz-Carrión, R.; Fuentes, J.; Redruello, Begoña; Guzmán, José M.; Río, María P. Martín del; Power, Deborah; Mancera, J. M.Prolactin (PRL), growth hormone (GH) and somatolactin (SL) expression was studied in gilthead sea bream (Sparus auratus) in response to several different stressors (salinity, food deprivation or stocking density). In the first experiment, specimens were acclimated during 100 days at three different environmental salinities: low salinity water (LSW, 6 ppt), brackish water (BW, 12 ppt) and seawater (SW, 38 ppt). Osmoregulatory parameters corresponded to those previously reported for this species under similar osmotic conditions. Pituitary PRL expression increased with decreasing environmental salinity, and was significantly different between SW- and LSW-acclimated fish. Pituitary GH expression was similar between SW- and BW-acclimated fish but decreased in LSW-acclimated specimens. Pituitary SL expression had a ‘‘U-shaped” relationship to environmental salinity with the lowest expression in BW-acclimated fish. In a second experiment SW-acclimated specimens were randomly assigned to one of four treatments and maintained for 14 days: (1) fed fish under low density (LD, 4 kg m!3); (2) fed fish under high density (HD, 70 kg m!3); (3) food deprived fish under LD; and (4) food deprived fish under HD. Plasma glucose and cortisol levels corresponded to those previously reported in S. auratus under similar experimental conditions. Pituitary PRL and SL expression increased in fish maintained under HD and decreased in food deprived fish. In conclusion, an effect of environmental salinity on pituitary PRL and GH expression has been demonstrated. In addition, crowding stress seems to interact with food deprivation in S. auratus and this is reflected by changes in pituitary PRL, GH and SL expression levels.