Dataset Information

Quantitative differences in rumen epithelium proteins in lambs fed wheat, perennial wheat or perennial wheat plus lucerne.

ABSTRACT: The rumen is a major the site of digestion and absorption of energy predominantly in the form of short chain fatty acids as well as amino acids and simple carbohydrates. The value of crops such as perennial wheat (PW) for grain and grazing compared to conventional wheat (W), or the addition of lucerne to PW (PWL) is still being determined. The research sought to determine if these diets representing high potassium (K): low sodium (Na) ratio (W, PW and PWL) was associated with changes in the membrane bound proteins that transport nutrients in the rumen epithelium (RE). Information about these proteins and the influence of different diets is scarce due to difficulties developing a procedure to examine them using quantitative proteomics. The research focuses on proteins extracted using an enzymatic procedure (Bond et al. 2019) from the rumen epithelium layer of the sheep’s’ forestomach. A membrane protein enriched fraction was then extracted and prepared for TMT-MS. The comprehensive assignment of proteins (>4000) to cellular location and quantification of differences in protein abundance will progress the annotation and understanding of protein functions in the ruminant genomes.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Ovis Aries

TISSUE(S): Rumen Epithelium, Epithelial Cell

DISEASE(S): Disease Free

SUBMITTER: Karthik Shantharam kamath

LAB HEAD: Dr Jude Bond

PROVIDER: PXD044135 | Pride | 2023-10-24

REPOSITORIES: Pride

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Publications

Quantitative Differences in Rumen Epithelium Proteins in Lambs Fed Wheat, Perennial Wheat, or Perennial Wheat plus Lucerne.

Bond Jude Jessie JJ Refshauge Gordon G Newell Matthew T MT Holman Benjamin W B BWB Wheeler David D Woodgate Serey S Kamath Karthik S KS Hayes Richard C RC

Proteomes 20230920 3

The value of crops such as perennial wheat (PW) for grain and grazing compared to conventional wheat (W), or the addition of lucerne to PW (PWL) is still being determined. This research sought to determine if these diets were associated with changes in the membranebound proteins that transport nutrients in the rumen epithelium (RE). Crossbred ewes (Poll Dorset × Merino) were fed W, PW, or PWL (50:50) fresh-cut forage <i>ad libitum</i> for 4 weeks. Average daily gain (ADG; <i>p</i> < 0.001) was h ...[more]

PMID: 37755706

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Project description:High levels of hepcidin, the main regulator of systemic iron metabolism leads to various diseases. Targeting hepcidin and lowering its concentration is a possible form of intervention in order to treat these diseases. High turnover rate of hepcidin is a major drawback of therapies directly targeting this peptide. We developed two monoclonal antibodies (mAbs) ABT-207 and h5F9-AM8 which inhibit hemojuvelin also known as repulsive guidance molecule c (RGMc) and downregulate hepcidin. After a single application of these antibodies hepcidin expression in liver and its concentration in serum were reduced. Serum iron increased for several weeks. The RGMc antibodies show a pronounced dose response relationship in rats with h5F9-AM8 having an IC50 (UIBC) of ~80 fold higher than ABT-207. When hepcidin levels were downregulated iron deposition in the liver was visible histologically one week post application. The anitbody-mediated iron deposition was not associated with any toxicologically relevant effect at the doses and timepoints evaluate. Iron depositions seen after 14 weekly treatments with ABT-207 were partially reversible in rats and in cynomolgus monkeys. Due to their long-lasting effects and excellent safety profile, both RGMc-blocking antibodies ABT-207 and h5F9-AM8 are favorable clinical candidates for diseases characterized by high serum hepcidin levels like anemia of chronic disease. Female Sprague-Dawley rats [Crl:CD®(SD)IGS BR], weighing ~200 g at study initiation were obtained from Charles River Laboratories, Inc. Rats were housed singly in ventilated, stainless steel, wire-bottom hanging cages, fed non-certified Rodent Chow and water ad libitum and acclimated for at least 5 days after arrival. Rats were randomly assigned to various treatment groups (5 rats/group) and were dosed once weekly by IV injection via tail vein with vehicle (30mM Histidine, 8% w/v Sucrose, pH6.0, + 0.02% Tween 80) or with 0.02, 0.2, or 20 mg/kg of RGMc antibody for a total of 4 doses. All rats were sacrificed under isoflurane anesthesia 24 hours after final dose. Liver and spleen waere flash frozen in liquid nitrogen and stored at 80°C until processing for gene expression profiling on the Affymetrix platform. total of 12 samples, 3 biological replicates (rats) from each of 4 treatments groups 1. vehicle control (30mM Histidine, 8% w/v Sucrose, pH6.0, + 0.02% Tween 80) 2. 0.02 mg/kg RGMC Ab 3. 0.2 mg/kg RGMC Ab 4. 20 mg/kg RGMC Ab; rats were dosed by IV injection via tail vein once weekly for 4 total doses, dose volume 2mL/kg/week

Dataset Information

Quantitative differences in rumen epithelium proteins in lambs fed wheat, perennial wheat or perennial wheat plus lucerne.

Publications

Quantitative Differences in Rumen Epithelium Proteins in Lambs Fed Wheat, Perennial Wheat, or Perennial Wheat plus Lucerne.

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