Myogenin (myog) encodes a highly conserved myogenic regulatory factor that is involved in terminal muscle differentiation. It has been shown in mammals that methylation of cytosines within the myog promoter plays a major role in regulating its transcription. In the present study, the Senegalese sole (Solea senegalensis) myog putative proximal promoter was identified and found to be highly conserved among teleosts. Therefore, it is plausible that it plays a similar role in controlling myog expression. Cytosine methylation of the myog promoter in skeletal muscle of Senegalese sole larvae undergoing metamorphosis was influenced by rearing temperature. A lower temperature (15°C) significantly increased myog promoter methylation in skeletal muscle, particularly at specific CpG sites, relatively to higher rearing temperatures (18 and 21°C). Myog transcription was downregulated at 15°C, whereas expression of dnmt1 and dnmt3b was upregulated, consistently with the higher myog methylation observed at this temperature. Rearing temperature also affected growth and fast muscle cellularity, producing larger fibers at 21°C. Taken together, our data provide the first evidence of an epigenetic mechanism that may be underlying the temperature-induced phenotypic plasticity of muscle growth in teleosts.
A study was undertaken to determine the dietary protein level for optimal growth performance and body composition of juvenile Senegalese sole. Five experimental extruded diets were formulated to contain increasing levels of protein [430, 480, 530, 570 and 600 g kg )1 dry matter (DM)] and a constant lipid level, ranging from 100 to 130 g kg )1 DM. Triplicate groups of 35 sole (initial body weight: 11.9 ± 0.5 g) were grown over 84 days in 60-L tanks supplied with recirculated seawater. Fish were fed by means of automatic feeders in eight meals per day. At the start and end of the trial, whole-body samples were withdrawn for proximate composition analysis. At the end of 84 days of experimental feeding, daily weight gain and specific growth rate in fish fed diets P43 and P48 were significantly lower than those found in fish fed higher protein level diets (P53, P57 and P59). Similarly, feed efficiency was also significantly lower in fish fed diet P43 than in fish fed all other dietary treatments. Sole juveniles fed lower protein level diets (P43 and P48) showed a significantly lower protein content than fish fed the higher dietary protein level treatments (P53, P57 and P60). Changes within the tested dietary protein levels did not affect significantly protein productive value or total nitrogen (N) losses in fish. However, daily N gain was significantly higher (P < 0.05) in fish fed diets P53 and P60 than in fish fed the lowest protein level diet (P43). Data from the present study indicate that diets for juvenile Senegalese sole should include at least 53% crude protein to maintain a good overall growth performance. Based on a second-order polynomial regression model, the daily crude protein requirement for maximum whole-body N gain as estimated here for Senegalese sole juveniles was 6.43 g kg )1 body weight day )1 which corresponds to a value of 1.03 g N intake kg )1 body weight day )1 . If the present data are expressed on a dietary crude protein concentration basis, the allowance for maximum protein accretion (N gain) would be met by a diet containing a crude protein level of 600 g kg )1 .
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