Acute heat stress and reduced nutrient intake alter intestinal proteomic profile and gene expression in pigs

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Abstract

Heat stress and reduced feed intake negatively affect intestinal integrity and barrier function. Our objective was to compare ileum protein profiles of pigs subjected to 12 hours of HS, thermal neutral ad libitum feed intake, or pair-fed to heat stress feed intake under thermal neutral conditions (pair-fed thermal neutral). 2D-Differential In Gel Electrophoresis and gene expression were performed. Relative abundance of 281 and 138 spots differed due to heat stress, compared to thermal neutral and pair-fed thermal neutral pigs, respectively. However, only 20 proteins were different due to feed intake (thermal neutral versus pair-fed thermal neutral). Heat stress increased mRNA expression of heat shock proteins and protein abundance of heat shock proteins 27, 70, 90-α and β were also increased. Heat stress reduced ileum abundance of several metabolic enzymes, many of which are involved in the glycolytic or TCA pathways, indicating a change in metabolic priorities. Stress response enzymes peroxiredoxin-1 and peptidyl-prolyl cis-trans isomerase A were decreased in pairfed thermal neutral and thermal neutral pigs compared to heat stress. Heat stress increased mRNA abundance markers of ileum hypoxia. Altogether, these data show that heat stress directly alters intestinal protein and mRNA profiles largely independent of reduced feed intake. These changes may be related to the reduced intestinal integrity associated with heat stress.

Figures

  • Table 1. Effects of 12 h of thermal neutral (TN; 21°C) ad libitum, pair-feeding in thermal neutral conditions (PFTN) or heat stress (HS; 37°C) ad libitum conditions on pig phenotypicmeasures.
  • Fig 1. A representative 2D-DIGE gel from the ileum showing identified proteins. A total of 45 μg of CyDye labeled protein (15 μg of each CyDye 2, 3, and 5) was loaded onto a 11 cm pH 3–10 IPG strip for the first dimension and the second dimension was run on a 12.5% SDS-PAGE gel. Proteins labeled with CyDye5 are shown.
  • Table 2. Differentially abundant proteins between thermal neutral (TN) and heat-stressed (HS) pigs at 12 h.
  • Table 3. Differentially abundant proteins between pair-fed thermal neutral (PFTN) and heat-stressed (HS) pigs at 12 h.
  • Fig 2. Detection of heat shock protein 70 (HSP 70) in ileum by the comparison of a (A) 2D-DIGE image (CyDye 5) and (B) western blot. 2D-DIGE performed using 15 μg of protein labeled with each CyDye loaded on an 11 cm pH 3–10 strip for the first dimension and the second dimension on a 12.5% SDS-PAGE gel (acrylamide: N,N0-bis-methylene acrylamide 100:1, 0.1% SDS, 0.05% TEMED, 0.05% ammonium persulfate, and 0.5 M Tris–HCL, pH 8.8). Western blot protocols were followed after loading 40 μg of protein on a 7 cm 3–10 pH strip and running the second dimension on a 12.5% SDS-PAGE gel.
  • Table 4. Effects of ad-libitum feed intake in thermal neutral conditions (TN; 21°C), ad-libitum feed intake in heat stress conditions (HS; 37°C), or pair-feeding in thermal neutral conditions (PFTN) on ileal mRNA abundance.
  • Table 5. Categorization and general function information for proteins identified as differentially expressed.

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APA

Pearce, S. C., Lonergan, S. M., Huff-Lonergan, E., Baumgard, L. H., & Gabler, N. K. (2015). Acute heat stress and reduced nutrient intake alter intestinal proteomic profile and gene expression in pigs. PLoS ONE, 10(11). https://doi.org/10.1371/journal.pone.0143099

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