iTRAQ-based quantitative proteomic analysis of alterations in the intestine of Hu sheep under weaning stress

Abstract

When Lambs are weaned off ewe’s milk, metabolic, structural, and functional changes often occur in the small intestine. Because information on the effects of weaning stress on the proteome of the intestine is limited, an animal model was established with eight pairs of twin lambs divided into artificially reared and ewe-reared groups, which was followed by proteome analysis using iTRAQ technology. Changes occurred in the morphology of the intestine and 5,338 proteins in three biological replicates with less than a 1.2% false discovery rate were identified and quantified. Among them, a subset of 389 proteins were screened as significantly up- (143) and down-regulated (246) in artificially reared compared with ewe-reared. According to Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, the differentially expressed proteins that were strongly down-regulated were enriched in immune system processes, biological adhesion, and metabolic processes. The up-regulated proteins were enriched in gene expression, cellular biosynthetic processes, ribosome and RNA binding in response to weaning stress. A series of proteins associated with intestine morphology and immune function were identified, and levels of the mRNAs encoding these proteins were analyzed by real-time quantitative reverse transcription PCR. The results of this study increased our understanding of the response of lambs weaned off ewe’s milk and helped to determine the mechanisms underlying weaning stress.

Effect of Age and Weaning on Growth Performance, Rumen Fermentation, and Serum Parameters in Lambs Fed Starter with Limited Ewe–Lamb Interaction

“…Similarly, this disease occurs most frequently within the two weeks after weaning and is characterized by a profuse diarrhea, dehydration, significant mortality, and loss of body weight of pigs [27]. Our previous study showed a sudden change in the feeding system due to early weaning resulted in changes of the intestinal morphology and functions [28], that may trigger reductions in digestive and absorptive capacities. Studies suggest that increasing weaning age reduces stress associated with this period and allows animals to have a more mature gastrointestinal tract and become increasingly familiar with solid feed during milk-period with an improvement in growth performance [29,30].…”

Transcriptome and iTRAQ-Based Proteome Reveal the Molecular Mechanism of Intestinal Injury Induced by Weaning Ewe's Milk in Lambs

“…The same animals were used according to our previous study ( 6 ). In summary, eight pairs of twin neonatal ram Hu lambs born on the same day with almost the same body weight (2.75 ± 0.2 kg) were randomly divided into the two groups.…”

“…A PPI network was drawn using Cytoscape and the cytoHubba application was used to identify the top 10 hub genes ( 19 , 20 ). The isobaric tags for relative and absolute quantification (iTRAQ) data of the same samples has been published in the previous study ( 6 ). In this integrative study, DEGs were identified by RNA-seq.…”

“…However, weaning is the most severe stimulating factor for the animals during early development, which is shown as the decreased feed intake, low nutrition absorption, aberrant mucosal immunity, and higher mortality ( 3 – 5 ). Young ruminants' intestinal histology and function would significantly change after the colonization of residential microorganisms, invasion of exogenous pathogens, and deletion of maternal antibodies ( 6 – 8 ), closely related to their growth, inflammatory response, and susceptibility to some diseases ( 5 ).…”

Identification of possible new salivary biomarkers of stress in sheep using a high-resolution quantitative proteomic technique

“…In veterinary science, recent investigations using these novel quantitative technologies to detect possible biomarkers of diagnostic and understand the prognosis of diseases in dogs (Martínez-Subiela et al, 2017;Franco-Martínez et al, 2018) have been reported. Also, studies performed in sheep using these techniques to evaluate the alterations in the intestine associated with under weaning stress using jejunum samples (Cui et al, 2018) or to consider age-related changes in the cerebrospinal fluid (Chen et al, 2018) have been published. However, to the best of our knowledge, no reports currently exist on the use of isobaric tagging technology in saliva samples to detect possible biomarkers of stress in sheep.…”