Browsing by Author "Geurden, Inge"
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- Dietary glucose stimulus at larval stage modifies the carbohydrate metabolic pathway in gilthead seabream (Sparus aurata) juveniles: An in vivo approach using C-14-starchPublication . Rocha, Filipa; Dias, Jorge; Geurden, Inge; Dinis, Maria Teresa; Panserat, Stephane; Engrola, S.The concept of nutritional programming was investigated in order to enhance the use of dietary carbohydrates in gilthead seabream juveniles. We assessed the long-term effects of high-glucose stimuli, exerted at the larval stage, on the growth performance, nutrient digestibility and metabolic utilization and gene expression of seabream juveniles, challenged with a high-carbohydrate intake. During early development, a group of larvae (control, CTRL) were kept under a rich-protein-lipid feeding regime whereas another group (GLU) was subjected to high-glucose stimuli, delivered intermittently over time. At juvenile stage, triplicate groups (IBW: 2.5 g) from each fish nutritional background were fed a high-protein (59.4%) low-carbohydrate (2.0%) diet before being subjected to a low-protein (43.0%) high-carbohydrate (33.0%) dietary challenge for 36-days. Fish from both treatments increased by 8-fold their initial body weight, but neither growth rate, feed intake, feed and protein efficiency, nutrient retention (except lipids) nor whole-body composition were affected (P (>) 0.05) by fish early nutritional history. Nutrient digestibility was also similar among both groups. The metabolic fate of C-14-starch and C-14-amino acids tracers was estimated; GLU juveniles showed higher absorption of starch-derived glucose in the gut, suggesting an enhanced digestion of carbohydrates, while amino acid use was not affected. Moreover, glucose was less used for de novo synthesis of hepatic proteins and muscle glycogen from GLU fish (P < 0.05). Our metabolic data suggests that the early glucose stimuli may alter carbohydrate utilization in seabream juveniles. (C) 2016 Elsevier Inc. All rights reserved.
- Glucose metabolism and gene expression in juvenile zebrafish (Danio rerio) challenged with a high carbohydrate diet: effects of an acute glucose stimulus during late embryonic lifePublication . Rocha, Filipa; Dias, Jorge; Engrola, S.; Gavaia, Paulo; Geurden, Inge; Dinis, Maria Teresa; Panserat, StephaneKnowledge on the role of early nutritional stimuli as triggers of metabolic pathways in fish is extremely scarce. The objective of the present study was to assess the long-term effects of glucose injection in the yolk (early stimulus) on carbohydrate metabolism and gene regulation in zebrafish juveniles challenged with a high-carbohydrate low-protein (HC) diet. Eggs were microinjected at 1 d post-fertilisation (dpf) with either glucose (2 M) or saline solutions. Up to 25 dpf, fish were fed a low-carbohydrate high-protein (LC) control diet, which was followed by a challenge with the HC diet. Survival and growth of 35 dpf juveniles were not affected by injection or the HC diet. Glucose stimulus induced some long-term metabolic changes in the juveniles, as shown by the altered expression of genes involved in glucose metabolism. On glycolysis, the expression levels of hexokinase 1 (HK1) and phosphofructokinase-6 (6PFK) were up-regulated in the visceral and muscle tissues, respectively, of juveniles exposed to the glucose stimulus, indicating a possible improvement in glucose oxidation. On gluconeogenesis, the inhibition of the expression levels of PEPCK in fish injected with glucose suggested lower production of hepatic glucose. Unexpectedly, fructose-1,6-bisphosphatase (FBP) expression was induced and 6PFK expression reduced by glucose stimulus, leaving the possibility of a specific regulation of the FBP-6PFK metabolic cycle. Glucose metabolism in juveniles was estimated using a [C-14]glucose tracer; fish previously exposed to the stimulus showed lower retention of [C-14]glucose in visceral tissue (but not in muscle tissue) and, accordingly, higher glucose catabolism, in comparison with the saline group. Globally, our data suggest that glucose stimulus at embryo stage has the potential to alter particular steps of glucose metabolism in zebrafish juveniles.
- Glucose overload in yolk has little effect on the long-term modulation of carbohydrate metabolic genes in zebrafish (Danio rerio)Publication . Rocha, Filipa; Dias, J.; Engrola, S.; Gavaia, Paulo J.; Geurden, Inge; Dinis, Maria Teresa; Panserat, S.The use of early nutritional stimuli to program metabolic pathways in fish is ill defined. Therefore, studies were undertaken with zebrafish to assess the effect of high glucose levels during the embryonic stage as a lifelong modulator of genes involved in carbohydrate metabolism. Genes related to carbohydrate metabolism were expressed at low levels at 0.2 and 1 day post-fertilization (dpf). However, from 4 dpf onwards there was a significant increase on expression of all genes, suggesting that all analysed pathways were active. By microinjection, we successfully enriched zebrafish egg yolk with glucose (a 43-fold increase of basal levels). Acute effects of glucose injection on gene expression were assessed in larvae up to 10 dpf, and the programming concept was evaluated in juveniles (41 dpf) challenged with a hyperglucidic diet. At 4 dpf, larvae from glucose-enriched eggs showed a downregulation of several genes related to glycolysis, glycogenolysis, lipogenesis and carbohydrate digestion in comparison with control (saline-injected) embryos. This inhibitory regulation was suppressed after 10 dpf. At the juvenile stage, and upon switching from a low to a high digestible carbohydrate diet, early glucose enrichment had no significant effect on most analysed genes. However, these same fish showed altered expression of the genes for cytosolic phosphoenolpyruvate carboxykinase, sodium-dependent glucose cotransporter 1 and glycogen synthase, suggesting changes to the glucose storage capacity in muscle and glucose production and transport in viscera. Overall, supplementation of egg yolk with high glucose levels had little effect on the long-term modulation of carbohydrate metabolic genes in zebrafish.
- High-glucose feeding of gilthead seabream (Sparus aurata) larvae: effects on molecular and metabolic pathwaysPublication . Rocha, Filipa; Dias, Jorge; Geurden, Inge; Dinis, Maria Teresa; Panserat, Stephane; Engrola, S.Nutritional programming has begun to arouse interest as a novel tool to alter specific metabolic pathways or functions in farmed animals. The aim of the present study was to explore the potential of early glucose stimuli to induce changes in nutrient metabolism of gilthead seabream. Nutritional conditioning was performed by delivering glucose-rich feed at three distinct recurrent periods of larval feeding regime: during first-feeding with rotifers (3 days after hatching, DAH) and mid-feeding with Artemia metanauplii (20DAH) and the beginning of inert diet feeding (30DAH), called the Recurrent treatment (REC). As opposed, the control treatment (CTRL) did not experience any glucose stimuli. At post-larval stage (from 50 to 60DAH), both treatments were challenged with a high-carbohydrate diet (50%). The immediate response to the early stimuli was assessed through gene expression of metabolic markers and by nutrient metabolism using [C-14] tracers. Each dietary stimulus induced metabolic changes on REC larvae, shown by altered expression of some genes, including those involved in glycolysis, and by a different pattern of glucose utilization. However, none of the molecular adaptations (except G6PDH gene) were persistent in the viscera and muscle of challenged post-larvae from REC group. In contrast, the glucose metabolism of challenged REC post-larvae revealed a shift towards a higher catabolism and lower glucose retention in tissues, compared to the CTRL group, suggesting an improvement of glucose oxidation pathways. In addition, the REC group showed a higher bio-conversion of glucose into lipids, indicating enhanced hepatic lipogenesis. The early stimuli did not affect the relative retention or use of amino acids or the growth and survival of challenged fish, up to 60DAH. In summary, although not substantiated at a molecular level, our data reveal that a recurrent high-glucose stimulus during larval stages affects the short-term modulation of pathways for glucose utilization in gilthead seabream. (C) 2015 Elsevier B.V. All rights reserved.
- Parental and early-feeding effects of dietary methionine in rainbow trout (Oncorhynchus mykiss)Publication . Fontagne-Dicharry, Stephanie; Alami-Durante, Helene; Aragão, Cláudia; Kaushik, Sadasivam J.; Geurden, IngeWe studied the effect of changes in dietary methionine (Met) supply in broodstock and first-feeding rainbow trout fry (offspring). Three plant-based diets differing in Met level (deficient, adequate or in excess of the established requirement) were fed to the broodstock (male and female) for 6 months prior to spawning (diets BD, BA and BE, respectively). The offspring from the parental Met-groups was then challenged in turn with the different Met fry-diets (FD, FA and FE, respectively) for 3 weeks from first-feeding. At spawning, females fed diet BD had significantly higher plasma total and LDL-cholesterol and slightly lower plasma triacylglycerol. Diet BD reduced female (but not male) growth, weight of spawn and egg size, but had no effect on relative fecundity. The free amino acid profile of oocytes was modified, with levels of Met and Cys correlating positively with the Met-levels of broodstock diets. SAM and SAH levels in oocytes followed the same pattern, as opposed to SAM/SAH ratio. At the swim-up stage, no significant effect of parental diet on fry weight was noted, whereas survival was the highest in fry from BE-broodstock. The subsequent 21-day fry feeding with different Met levels highly affected the daily growth index with a significant interaction between the parental-diet and fry-diet effects. The expression of a number of genes regulating sulfur amino acid metabolism was modified either directly by the dietary Met supply in both broodstock liver and in whole fry (e.g. BHMT1, GR, GST pi, MsrA1) or indirectly by the parental Met intakes as seen in the swim-up fry (e.g. BHMT1, MTR, GST pi, MsrA1). Importantly, long-lasting parental effects linked to broodstock Met-intake were seen in the fry, 21-days after first-feeding and irrespective of the fry diet (CTH, MsrA1, MsrB2, SOD2). Similarly, parental effects were noted on the gene expression of both NPY and POMC feeding peptides in fry prior to exogenous feeding which persisted for POMC in the 21-day fry. Parental effects were also demonstrated on the key myogenic gene Myog, on fMHC and GDH in swim-up fry, which persisted for GDH in 21-day fry. In summary, our results demonstrate that dietary Met levels of rainbow trout broodstock affect various traits in the offspring, some of which persisted during the first weeks of exogenous feeding. Further studies need to evaluate the long-term persistence of the parental effects over time and to elucidate the mechanisms, whether epigenetic or not. Statement of relevance: Determining the multiple effects of dietary methionine levels on reproductive, growth performance and metabolism in offspring will help improve formulations of low fish meal feeds for rainbow trout at sensitive life cycle stages. (C) 2016 Elsevier B.V. All rights reserved.
- Programming the metabolic gene expression in zebrafish juveniles (Danio rerio) by supra-physiological glucose level during embryogenesisPublication . Rocha, F.; Dias, J.; Engrola, S.; Gavaia, Paulo J.; Geurden, Inge; Dinis, Maria Teresa; Panserat, S.Using zebrafish as a model organism, assess the effect of high glucose levels during early embryonic stages as a lifelong moderator of metabolic pathways.