Abstract
Methionine (Met) is an important building block and metabolite for protein biosynthesis. However, the mechanism behind its absorption in the fish gut has not been elucidated. Here, we describe the fundamental properties of Met transport along trout gut at µmol/L and mmol/L concentration. Both electrogenic and unidirectional DL-[(14) C]Met flux were employed to characterize Met transporters in Ussing chambers. Exploiting the differences in gene expression between diploid (2N) and triploid (3N) and intestinal segment as tools, allowed the association between gene and methionine transport. Specifically, three intestinal segments including pyloric caeca (PC), midgut (MG), and hindgut (HG) were assessed. Results at 0-150 µmol/L concentration demonstrated that the DL-Met was most likely transported by apical transporter ASCT2 (SLC1A5) and recycled by basolateral transporter y(+) LAT1 (SLC7A7) due to five lines of observation: (1) lack of Na(+) -independent kinetics, (2) low expression of B(0) AT2-like gene, (3) Na(+) -dependent, high-affinity (K(m) , µmol/L ranges) kinetics in DL-[(14) C]Met flux, (4) association mRNA expression with the high-affinity kinetics and (5) electrogenic currents induced by Met. Results at 0.2-20 mmol/L concentration suggested that the DL-Met transport is likely transported by B(0) AT1-like (SLC6A19-like) based on gene expression, Na(+) -dependence and low-affinity kinetics (K(m) , mmol/L ranges). Similarly, genomic and gene expression analysis suggest that the basolateral exit of methionine was primarily through LAT4-like transporter (SLC43A2-like). Conclusively, DL-Met uptake in trout gut was most likely governed by Na(+) -dependent apical transporters ASCT2 and B(0) AT1-like and released through basolateral LAT4-like, with some recycling through y(+) LAT1. A comparatively simpler model than that previously described in mammals.