Multi-tissue transcriptomic analysis reveals that L-methionine supplementation maintains the physiological homeostasis of broiler chickens than D-methionine under acute heat stress

PLoS One. 2021 Jan 27;16(1):e0246063. doi: 10.1371/journal.pone.0246063. eCollection 2021.

Abstract

The objective of this study was to compare the effects of supplementation with two methionine isoforms, L-methionine (L-Met) or D-methionine (D-Met), on transcriptome expression in broiler chickens under acute heat stress. A total of 240 one-day-old chicks were randomly assigned to one of four treatments in a 2 × 2 factorial arrangement: thermo-neutral vs. acute heat-stress and L-Met vs. D-Met supplementation. On day 14, the heat-stressed group was exposed to 32°C for 5 h, while the others remained at 25°C. Six chicks were randomly selected per treatment and total RNA was isolated from whole blood, ileum, and liver tissues. Two RNA samples from each tissue of each treatment group were randomly selected and pooled in equal amounts. A total of 1.87 billion raw reads obtained from 36 samples (four treatments × three tissues × three composited replicates) were mapped to the reference genome build (Gallus_gallus-5.0) and used to identify differentially expressed genes (DEGs) using DESeq2. Functional enrichment of DEGs was tested using DAVID. Comparing the two isoforms of supplemented methionine, two, three, and ten genes were differentially expressed (> 1 or < -1 log2 fold change) in whole blood, ileum, and liver, respectively. A total of 38, 71, and 16 genes were differentially expressed in response to the interaction between heat stress and Met isoforms in the blood, ileum, and liver, respectively. Three-tissue-specific DEGs were functionally enriched for regulation of cholesterol homeostasis and metabolism, glucose metabolism, and vascular patterning. Chicks fed with L-Met had lower immune (e.g., IL4I1 and SERPINI1) and intestinal angiogenic responses (e.g., FLT1 and FGD5), and stable glucose and lipid metabolism (e.g., PCK1 and LDLR) under heat stress conditions. In conclusion, unlike D-Met, L-Met supplementation seems to help maintain physiological homeostasis and enhances cellular defense systems against external stresses like high environmental temperature.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animal Feed / analysis
  • Animal Husbandry
  • Animals
  • Chickens / genetics*
  • Chickens / physiology
  • Dietary Supplements / analysis
  • Heat Stress Disorders / genetics
  • Heat Stress Disorders / prevention & control
  • Heat Stress Disorders / veterinary
  • Heat-Shock Response / drug effects*
  • Methionine / pharmacology*
  • Transcriptome / drug effects*

Substances

  • Methionine

Grants and funding

This work performed by the Chungnam National University was financially supported by the CJ Cheiljedang Co. Ltd. (Seoul, Republic of Korea). The funds were used for the purchase of experimental animals and materials, and RNA-sequencing analysis costs. The L-methionine product used in this study is a commercial product invented by CJ Cheiljedang Co. The funder provided support in the form of research material and analysis cost for authors [M.L., H.P., J.M.H., and S.S.], but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. H.J.C. is an employee of CJ Cheiljedang Co., but was not the inventor of the product used in the research. The specific roles of these authors are articulated in the ‘author contributions’ section.