Abstract

The effects of dietary methionine (Met) supply on the postprandial pattern of plasma free amino acids and the differential expression of several genes associated with a number of sulfur amino acid and protein turnover pathways in the liver of juvenile barramundi (Lates calcarifer) was investigated. At the conclusion of a 49-day growth trial assessing the requirement for dietary Met, three treatments were selected (with deficient (DEF; 8.6 g kg−1), adequate (ADQ; 14.9 g kg−1) and excessive (EXC; 21.4 g kg−1)) levels of dietary Met, based on their respective growth responses. A peak occurred in plasma free Met at 2 h post-feeding in fish fed the DEF and ADQ diets and at 4 h post-feeding in fish in the EXC treatment. Liver samples collected at these timepoints, as well as those taken as a pre-feeding control, were analyzed for expression of genes involved in Met turnover (CGL, MAT-1, MAT-2a) and taurine biosynthetic pathways (CSAD, ADO, CDO), target of rapamycin inhibition (Redd-1), the somatotropic axis (GHR-II, IGFI, IGF-II) and protein turnover pathways (MUL-1, ZFAND-5). Markers of sulfur amino acid turnover were more significantly affected by time after feeding than by dietary Met level, suggesting production of these enzymes may be primarily regulated by the consumption of feed or protein, rather than by the dietary composition. Further, metabolised Met appeared likely to have been directed through S-Adenosylmethionine (SAM) dependent pathways, rather than converted to Cys, which may have contributed to the observed growth response. Both genes influencing the conversion of Met to SAM appear to be active at this lifestage in barramundi. Previously described markers of proteolytic pathways appear to be conserved in this species and we have confirmed that ZFAND-5 is a reliable biomarker of this process in barramundi. A number of important genes were investigated for the first time in this species and shown to be nutritionally regulated.

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