Differential expression of proteins involved in energy production along the crypt-villus axis in early-weaning pig small intestine

Abstract

Weaning of piglets reflects intestinal dysfunction and atrophy and affected the physiological state of enterocytes. However, few studies have defined physiological state of enterocytes along the crypt-villus axis in early-weaning piglets. A total of 16 piglets from 8 litters were used in the experiment. One group of piglets was nursed by sows until age 21 days, and another group was weaned at age 14 days and then fed creep feed instead of breast milk for 7 days. Piglets were killed at 21 days, and the jejunum segments were dissected. After sequential isolation of jejunum epithelial cells along the crypt-villus axis, their proteins were analyzed through the isobaric tags for relative and absolute quantification, and proteins involved in the mammalian target of rapamycin signaling pathway and proliferating cell nuclear antigen abundances in jejunal epithelial cells of weaning or suckling group were determined by Western blotting. The differential proteins in three cell fractions were identified and analyzed. The results showed that proteins involved in the tricarboxylic acid cycle, ␤-oxidation, and the glycolysis pathway were significantly downregulated in the upper and middle villus of the early-weaned group. However, proteins involved in glycolysis were significantly upregulated in crypt cells. In addition, Western blot analysis showed that the expression of mammalian target of rapamycin pathway-related proteins was decreased (P ＜ 0.05) in the earlyweaned group. The present results showed that early-weaning differentially affect the expression of proteins involved in energy production of enterocytes along the jejunal crypt-villus axis.weaning; enterocyte; crypt-villus axis; piglet EARLY WEANING (14-to 21-day weaning age) of piglets is stressful and is characterized by poor growth, decreased feed intake, and diarrhea, because they are abruptly forced to combined stressors, including maternal and littermate separations, changes in diet, and establishment of a new social hierarchy (30, 31). Early weaning stress results in increased intestinal dysfunction and atrophy, impaired mucosal barrier function, and an imbalance of intestinal microbiota (13, 30). Inadequate food intake immediately after weaning and insufficient energy intake may contribute to the changes in intestinal morphology (25,35).The intestinal epithelium is characterized by rapidly proliferating cells organized into structures called crypts, which invaginate into the underlying mesenchyme, and villi, which project into the lumen (6). Furthermore, epithelial cells undergo functional and morphological differentiation during migration along the crypt-villus axis from the crypt to the tip of the villus (22, 43). Intestinal epithelial cell proliferation, differentiation, and apoptosis play important roles in intestinal development, maintenance, and recovery from tissue damage (44). In addition, epithelial cells in the pig intestine have notably high energy demands due to the rapid renewal of the epithelium (within few days) (2, 36). Previous studies showed...

Correlations of genes expression in PPAR signalling pathway with porcine meat quality traits

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A YY1-dependent increase in aerobic metabolism is indispensable for intestinal organogenesis

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Mammalian Target of Rapamycin Signaling Pathway Changes with Intestinal Epithelial Cells Renewal Along Crypt-Villus Axis

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“…The isolated cells were immediately frozen in liquid nitrogen and then stored at -80°C until analysis. The efficiency of fractionation cell was validated by measuring alkaline phosphatase and PCNA as previously described [12]. The activity of alkaline phosphatase was determined using an enzyme assay kit, according to the manufacturer's protocol (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) [13].…”