Project description:A more in-depth exploration of gut functional aspects may be interesting in order to provide hints for action (e.g. dietary strategies) to favor gut balance maintenance (Sinha et al., 2017), given the important role of the intestine in development of possible metabolic diseases. A careful survey on the differential gene expression may help to scouting new interesting functions and identify potential markers for testing various experimental factors. The transcriptomes of the jejunum and cecum mucosae of 19 broiler chickens were compared. At slaughter age (day 42), on 38 birds, selected with a homogeneous body weight, jejunum and cecum mucosae were collected by gently scraping after tissues rinsing in PSB to remove residues of digesta, and immediately frozen in liquid nitrogen and then stored at -80°C. From both tissues, total RNA was extracted using GeneJET RNA Purification Kit (Thermo Scientific)
Project description:Adding lysolecithin to feed has reportedly improved the performance of broiler chickens. Lysolecithin is generated by phospholipase catalyzed hydrolysis of lecithin. The enzymatic reaction converts phospholipids into lysophospholipids, with lysophosphatidylcholine (LPC) the primary product. Here we compared supplementation with a commercial lysolecithin (Lysoforte(R) Kemin Industries, Inc., Des Moines, IA) with comparable levels of purified LPC for effects on broilers. Despite no differences in weight gain during the starter period, we discovered a significant increase in average villus length in the jejunum with lysolecithin, but not with LPC. High-throughput gene expression microarray analyses revealed many more genes were regulated in the epithelium of jejunum by lysolecithin compared to LPC. The most upregulated genes and pathways were for collagen, extracellular matrix and integrins. Staining sections of jejunum with Sirius Red confirmed the increased deposition of collagen fibrils in villi of broilers fed lysolecithin but not LPC. Thus, lysolecithin elicits gene expression in the intestinal epithelium leading to enhanced collagen deposition and villus length. LPC alone as a supplement does not mimic these responses. Feed supplementation with lysolecithin triggers changes in the intestinal epithelium with the potential to improve overall gut health and performance.
Project description:The study proposed an exploratory functional analysis on differential gene expression of the jejunum and of cecum in chickens. For this study, 150 Ross 308 male chickens were randomly allotted in six pens (25 birds/pen) and fed the same commercial diet. From 19 birds of 42 days of age, jejunum and cecum mucosae were collected for RNA extraction for transcriptome microarray analysis. Differentially expressed genes (DEGs) submitted to DAVID (Database for Annotation, Visualization, and Integrated Discovery) and Gene Set Enrichment Analysis (GSEA) software evidenced enriched gene clusters for biological functions differentiated in the tissues. DAVID analysis in the jejunum showed enriched annotations for cell membrane integral components, PPAR (peroxisome proliferator-activated receptor) signaling pathway, and peroxisome and lipid metabolism, and showed DEGs for gluconeogenesis, not previously reported in chicken jejunum. The cecum showed enriched annotations for disulfide bond category, cysteine and methionine metabolism, glycoprotein category, cell cycle, and extracellular matrix (ECM). GSEA analysis in the jejunum showed peroxisome and PPAR signaling pathway-related gene sets, as found with DAVID, and gene sets for immune regulation, tryptophan and histidine metabolism, and renin-angiotensin system, like in mammals. The cecum showed cell cycle and regulation processes, as well as ECM receptor interaction and focal adhesion-related gene sets. Typical intestinal functions specific for the gut site and interesting functional genes groups emerged, revealing tissue-related key aspects which future studies might take advantage of.