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RNA-seq analysis of the kidneys of broiler chickens fed diets containing different concentrations of calcium.

ABSTRACT: Calcium (Ca) is required for normal growth and is involved in cellular physiology, signal transduction, and bone mineralization. In humans, inadequate Ca intake causes hypocalcaemia, and excessive Ca intake causes hypercalcemia. In chicken, Ca is also required for body weight gain and eggshell formation. However, transcriptomic responses to low/high Ca intake, and mechanisms affecting body weight have not been explored. In this study, we performed comparative RNA sequencing (RNA-seq) using the kidney of broiler chickens fed diets containing 0.8, 1.0, and 1.2% Ca. Annotation of RNA-seq data revealed a significant number of differentially expressed genes (DEGs) in the kidney via pairwise comparison using Cufflinks and edgeR. Using edgeR, we identified 12 DEGs; seven overlapped with those found by cufflinks. Seven DEGs were validated by real-time quantitative-PCR (qRT-PCR) in Ca-supplemented kidneys, and the results correlated with the RNA-seq data. DEGs identified by cufflinks/edgeR were subjected to pathway enrichment, protein/protein interaction, and co-occurrence analyses to determine their involvement in disease. The National Research Council (NRC) recommended Ca intake for 21-day post-hatch broilers is about 1.0%. Our findings suggest that higher-than-recommended Ca intake (1.2%) could reduce body weight gain in broilers, and that affected DEGs are related to stress-induced diseases, such as hypertension.

SUBMITTER: Park W

PROVIDER: S-EPMC5603577 | biostudies-literature | 2017 Sep

REPOSITORIES: biostudies-literature

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RNA-seq analysis of the kidneys of broiler chickens fed diets containing different concentrations of calcium.

Park Woncheoul W Rengaraj Deivendran D Kil Dong-Yong DY Kim Heebal H Lee Hak-Kyo HK Song Ki-Duk KD

Scientific reports 20170918 1

Calcium (Ca) is required for normal growth and is involved in cellular physiology, signal transduction, and bone mineralization. In humans, inadequate Ca intake causes hypocalcaemia, and excessive Ca intake causes hypercalcemia. In chicken, Ca is also required for body weight gain and eggshell formation. However, transcriptomic responses to low/high Ca intake, and mechanisms affecting body weight have not been explored. In this study, we performed comparative RNA sequencing (RNA-seq) using the k ...[more]

PMID: 28924246

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Project description:Trypsin inhibitors (TI) resident in soybeans affects protein utilization. While heat treatment influences residual TI, it simultaneously affects the structure and solubility of the soybean proteins and confounds any response to exogenous proteases. Using purified TI, the effect of exogenous protease to TI can be dissociated from changes in the soybean protein. Thus, the current study was designed to evaluate the growth performance and protein utilization responses of broiler chickens to purified TI and exogenous protease. Soybean meal (SBM) was preanalyzed for basal TI (2,996 TIU/g SBM), formulated into nutritionally adequate experimental diets to contain 1,033 TIU/g diet, and purified TI was added at 9,000 TIU/g diet. A total of 320 Cobb-500 broiler chicks were allocated to 4 diets, each with 8 replicate cages and 10 birds per replicate. The experimental diets were arranged as a 2 × 2 factorial with factors being dietary TI (1,033 or 10,033 TIU/g) and exogenous protease (0 or 15,000 PROT/kg). On day 7, 14, and 21 posthatching, protease supplementation improved the BW gain (P < 0.01) and gain to feed ratio (P < 0.05) of birds. On day 14 and 21 posthatching, the relative weight of pancreas increased (P < 0.05) with added TI but was reduced (P < 0.001) with protease supplementation. Apparent ileal digestibility of all amino acids, except methionine, decreased (P < 0.001) with added TI but increased (P < 0.05) with protease supplementation. Jejunal MUC-2 was downregulated (P < 0.01) and SCL7A-2 was upregulated (P < 0.05) by protease supplementation. Duodenal trypsin and chymotrypsin activities reduced (P < 0.05) with added TI but increased (P < 0.01) with protease supplementation. Exogenous protease produced longer villi (P < 0.05) and deeper crypts (P < 0.01) in the jejunal tissue. In conclusion, dietary addition of purified TI negatively affects nutrient utilization by broiler chickens. Furthermore, the study showed that the efficacy of the exogenous protease might be independent of dietary TI concentration.

Project description:An experiment was conducted to determine the digestible calcium (Ca) and digestible phosphorous (P) requirements of 10-day-old broiler chickens. Fifteen corn-soybean meal-based diets containing 3.3, 3.9, 4.4, 5.0, and 5.5 g/kg standardized ileal digestible (SID) Ca and 4.0, 5.0, and 6.0 g/kg SID P was fed to broilers from d 1 to 10. Each experimental diet was randomly allocated to 6 replicate cages (12 birds per cage). Body weight and feed intake were recorded at the start and end of the experiment and the feed conversion ratio was calculated. On d 10, birds were euthanized to collect ileal digesta, toes and tibia for the determination of digestible Ca and P, toe ash concentration and the concentrations of ash, Ca, and P in tibia. Titanium dioxide (5 g/kg) was included in all diets as an indigestible indicator for apparent ileal digestibility measurements. Total excreta were collected from d 1 to 10 for the measurement of total tract retention of Ca and P. Fixed effects of the experiment were dietary concentrations of SID Ca and SID P and their interaction. If the interaction or main effects were significant (P < 0.05), the parameter estimates for second-order response surface model were determined using General Linear Model procedure of SAS software. The growth performance, bone mineralization and mineral utilization of broiler starters were found to be optimized at 5 g/kg SID P concentration. Required SID Ca for maximum weight gain and bone mineralization was determined to be 3.32 and 4.36 to 4.78 g/kg, respectively, at 5 g/kg SID P concentration, which correspond to SID Ca to SID P ratios of 0.66 and 0.87 to 0.96, respectively. The estimated SID Ca requirement for weight gain is lower than the current Ca recommendation (9.6 g/kg total Ca or 4.4 g/kg SID Ca) for broiler starters. However, bone mineralization is maximized around the current total Ca recommendation at 8.9 to 9.8 g/kg (4.36-4.78 g/kg SID Ca) and indicates that bone mineralization requires more Ca than growth performance.

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RNA-seq analysis of the kidneys of broiler chickens fed diets containing different concentrations of calcium.

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RNA-seq analysis of the kidneys of broiler chickens fed diets containing different concentrations of calcium.

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