SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine
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. Bo...
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| Format: | Article |
| Language: | English |
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Wiley
2019-11-01
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| Series: | Physiological Reports |
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| Online Access: | https://doi.org/10.14814/phy2.14274 |
| _version_ | 1818302238183915520 |
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| author | Van P. T. H. To Karthik Masagounder Matthew E. Loewen |
| author_facet | Van P. T. H. To Karthik Masagounder Matthew E. Loewen |
| author_sort | Van P. T. H. To |
| collection | DOAJ |
| description | 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‐[14C]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 B0AT2‐like gene, (3) Na+‐dependent, high‐affinity (Km, µmol/L ranges) kinetics in DL‐[14C]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 B0AT1‐like (SLC6A19‐like) based on gene expression, Na+‐dependence and low‐affinity kinetics (Km, 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 B0AT1‐like and released through basolateral LAT4‐like, with some recycling through y+LAT1. A comparatively simpler model than that previously described in mammals. |
| first_indexed | 2024-12-13T05:35:44Z |
| format | Article |
| id | doaj.art-9184e405b2db40fbaf7ebf1cda15860c |
| institution | Directory Open Access Journal |
| issn | 2051-817X |
| language | English |
| last_indexed | 2024-12-13T05:35:44Z |
| publishDate | 2019-11-01 |
| publisher | Wiley |
| record_format | Article |
| series | Physiological Reports |
| spelling | doaj.art-9184e405b2db40fbaf7ebf1cda15860c2022-12-21T23:57:56ZengWileyPhysiological Reports2051-817X2019-11-01721n/an/a10.14814/phy2.14274SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestineVan P. T. H. To0Karthik Masagounder1Matthew E. Loewen2Veterinary Biomedical Sciences University of Saskatchewan Saskatoon Saskatchewan CanadaEvonik Nutrition & Care GmbH Rodenbacher Chaussee Hanau GermanyVeterinary Biomedical Sciences University of Saskatchewan Saskatoon Saskatchewan CanadaAbstract 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‐[14C]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 B0AT2‐like gene, (3) Na+‐dependent, high‐affinity (Km, µmol/L ranges) kinetics in DL‐[14C]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 B0AT1‐like (SLC6A19‐like) based on gene expression, Na+‐dependence and low‐affinity kinetics (Km, 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 B0AT1‐like and released through basolateral LAT4‐like, with some recycling through y+LAT1. A comparatively simpler model than that previously described in mammals.https://doi.org/10.14814/phy2.14274Electrogenicintestinemethionineradioisotope fluxrainbow trouttransport |
| spellingShingle | Van P. T. H. To Karthik Masagounder Matthew E. Loewen SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine Physiological Reports Electrogenic intestine methionine radioisotope flux rainbow trout transport |
| title | SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine |
| title_full | SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine |
| title_fullStr | SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine |
| title_full_unstemmed | SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine |
| title_short | SLC transporters ASCT2, B0AT1‐like, y+LAT1, and LAT4‐like associate with methionine electrogenic and radio‐isotope flux kinetics in rainbow trout intestine |
| title_sort | slc transporters asct2 b0at1 like y lat1 and lat4 like associate with methionine electrogenic and radio isotope flux kinetics in rainbow trout intestine |
| topic | Electrogenic intestine methionine radioisotope flux rainbow trout transport |
| url | https://doi.org/10.14814/phy2.14274 |
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