Expression of proposed methionine transporters along the gastrointestinal tract of pigs and their regulation by dietary methionine sources

Stella Romanet; Jörg R Aschenbach; Robert Pieper; Jürgen Zentek; John K Htoo; Rose A Whelan; Lucia Mastrototaro

doi:10.1186/s12263-021-00694-4

Expression of proposed methionine transporters along the gastrointestinal tract of pigs and their regulation by dietary methionine sources

Genes Nutr. 2021 Sep 6;16(1):14. doi: 10.1186/s12263-021-00694-4.

Authors

Stella Romanet¹, Jörg R Aschenbach², Robert Pieper³, Jürgen Zentek³, John K Htoo⁴, Rose A Whelan⁴, Lucia Mastrototaro¹

Affiliations

¹ Institute of Veterinary Physiology, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany.
² Institute of Veterinary Physiology, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany. joerg.aschenbach@fu-berlin.de.
³ Institute of Animal Nutrition, Freie Universität Berlin, Berlin, Germany.
⁴ Evonik Operations GmbH, Animal Nutrition Services, Hanau-Wolfgang, Germany.

Abstract

Background: Given the key role of methionine (Met) in biological processes like protein translation, methylation, and antioxidant defense, inadequate Met supply can limit performance. This study investigated the effect of different dietary Met sources on the expression profile of various Met transporters along the gastrointestinal tract (GIT) of pigs.

Methods: A total of 27 pigs received a diet supplemented with 0.21% DL-Met, 0.21% L-Met, or 0.31% DL-2-hydroxy-4-(methylthio)butanoic acid (DL-HMTBA). Changes in mRNA expression of B⁰AT1, ATB^0,+, rBAT, ASCT2, IMINO, LAT4, y⁺LAT1, LAT2, and SNAT2 were evaluated in the oral mucosa, cardia, fundus, pylorus, duodenum, proximal jejunum, middle jejunum, ileum, cecum, proximal colon, and distal colon, complemented by protein expression analysis of B⁰AT1, ASCT2, LAT2, and LAT4.

Results: Expression of all investigated transcripts differed significantly along the GIT. B⁰AT1, rBAT, y⁺LAT1, LAT2, and LAT4 showed strongest mRNA expression in small intestinal segments. ASCT2, IMINO, and SNAT2 were similarly expressed along the small and large intestines but expression differed in the oral mucosa and stomach. ATB^0,+ showed highest mRNA expression in large intestinal tissues, cardia, and pylorus. In pigs fed DL-Met, mRNA expression of ASCT2 was higher than in pigs fed DL-HMTBA in small intestinal tissues and mRNA expression of IMINO was lower than in pigs fed L-Met in large intestinal tissues. Dietary DL-HMTBA induced a stronger mRNA expression of basolateral uptake systems either in the small (LAT2) or large (y⁺LAT1) intestine. Protein expression of B⁰AT1 was higher in the middle jejunum and ileum in pigs fed DL-Met when compared with the other Met supplements. LAT4 expression was higher in pigs fed DL-HMTBA when compared with DL-Met (small intestine) and L-Met (small intestine, oral mucosa, and stomach).

Conclusion: A high expression of several Met transporters in small intestinal segments underlines the primary role of these segments in amino acid absorption; however, some Met transporters show high transcript and protein levels also in large intestine, oral mucosa, and stomach. A diet containing DL-Met has potential to increase apical Met transport in the small intestine, whereas a diet containing DL-HMTBA has potential to increase basolateral Met transport in the small intestine and, partly, other gastrointestinal tissues.

Keywords: Intestine; Longitudinal heterogeneity of gene expression; Methionine transport; Stomach; Western blot; qRT-PCR.