DNA methylation of the promoter region of bnip3 and bnip3l genes induced by metabolic programming.

BACKGROUND: Environmental changes of biotic or abiotic nature during critical periods of early development may exert a profound influence on physiological functions later in life. This process, named developmental programming can also be driven through parental nutrition. At molecular level, epigenetic modifications are the most likely candidate for persistent modulation of genes expression in later life. RESULTS: In order to investigate epigenetic modifications induced by programming in rainbow trout, we focused on bnip3 and bnip3l paralogous genes known to be sensitive to environmental changes but also regulated by epigenetic modifications. Two specific stimuli were used: (i) early acute hypoxia applied at embryo stage and (ii) broodstock and fry methionine deficient diet, considering methionine as one of the main methyl-group donor needed for DNA methylation. We observed a programming effect of hypoxia with an increase of bnip3a and the four paralogs of bnip3l expression level in fry. In addition, parental methionine nutrition was correlated to bnip3a and bnip3lb1 expression showing evidence for early fry programming. We highlighted that both stimuli modified DNA methylation levels at some specific loci of bnip3a and bnip3lb1. CONCLUSION: Overall, these data demonstrate that methionine level and hypoxia stimulus can be of critical importance in metabolic programming. Both stimuli affected DNA methylation of specific loci, among them, an interesting CpG site have been identified, namely - 884 bp site of bnip3a, and may be positively related with mRNA levels.

Rainbow trout prefer diets rich in omega-3 long chain polyunsaturated fatty acids DHA and EPA.

The control of feed intake in fish in aquaculture requires the development of new techniques to improve diet composition, feed conversion efficiency and growth. The aim must be sustainability and an effective use of resources. The effect of replacing traditional aqua-feed ingredients (fishmeal and fish oil) by a 100% plant-based diet is known to drastically decrease fish performance (survival and growth). The present study examined the feed preference of rainbow trout Oncorhynchus mykiss for three diets containing distinct levels of omega-3 long chain polyunsaturated fatty acids (ω-3 LCPUFA): eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) (0% for low, 5% for medium and 20% for high, total fatty acid content). Feed preference values for each group (low v. medium ω-3 diets, medium v. high ω-3 diets and low v. high ω-3 diets) were observed using two self-feeders positioned at opposite sides of the tank. The hypothesis was that the decrease of fish growth and survival rate of fish fed with 100% plant-based diet could be explained by the absence of ω-3 LCPUFA relating to decrease of food intake. This could explain the tasting role of ω-3 LCPUFA in the feeding behavior of rainbow trout (which reflects the motivation to consume feed). The results showed that rainbow trout could discriminate between the diets containing different level of ω-3 LCPUFA even if unable to differentiate between level of 5% (no preference observed in low v. medium ω-3 diets). Overall they had a preference for diet high in ω-3 LCPUFA: 59.5% preference for high ω-3 diet in high v. low ω-3 diets, and 75.6% preference for high ω-3 diet in medium v. high ω-3 diets respectively. This preference was repeated after 21 days and for a further 21 days when the feeds were exchanged between the two self- feeders in each tank: 63.3% preference for high ω-3 diet in high v. low ω-3 diets, and 69,5% preference for high ω-3 diet in medium v. high ω-3 diets respectively. The tests also indicated a difference in the extent of food waste of each of the three diets revealed by uneaten pellets after feed demands. During two periods of test, high ω-3 diet was the most appreciated, the least wasted and the most eaten (all choice groups) whereas the most uneaten feed remained the least appreciated diet in three choices diets (low ω-3 diet in low v. medium ω-3 diets, medium in medium v. high ω-3 diets and low in low v. high ω-3). In conclusion, this study highlighted the influence of ω-3 LCPUFA in the feeding behavior of juvenile rainbow trout, levels of ω-3 LCPUFA drove dietary choices in the fish.

Amino acids downregulate the expression of several autophagy-related genes in rainbow trout myoblasts.

Many fish species experience long periods of fasting often associated with seasonal reductions in water temperature and prey availability or spawning migrations. During periods of nutrient restriction, changes in metabolism occur to provide cellular energy via catabolic processes. Muscle is particularly affected by prolonged fasting as proteins of this tissue act as a major energy source. However, the molecular components involved in muscle protein degradation as well as the regulatory networks that control their function are still incompletely defined in fish. The present work aimed to characterize the response of the autophagy-lysosomal degradative pathway to nutrient and serum availability in primary culture of rainbow trout myoblasts. In this aim, 4-day-old cells were incubated in a serum and amino acid-rich medium (complete medium), a serum and amino acid-deprived medium (minimal medium) or a minimal medium plus amino acids, and both the transcription-independent short-term response and the transcription-dependent long-term response of the autophagy-lysosomal degradative pathway were analyzed. We report that serum and amino acids withdrawal is accompanied by a rapid increase of autophagosome formation but also by a slower induction of the expression of several autophagy-related genes (LC3B, gabarapl1, atg4b). We also showed that this latter response is controlled by amino acid (AA) availability and that both TOR-dependent and TOR-independent pathways are involved in this effect. Together these results suggest an important role for AA released by muscle proteolysis during the fasting period in regulating the subtle balance between using proteins as disposable furniture to provide energy, and conserving muscle through protein sparing.