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Prednisolone-induced predisposition to femoral head separation and the accompanying plasma protein changes in chickens.

Packialakshmi B, Liyanage R, Lay J, Okimoto R, Rath N - Biomark Insights (2015)

Bottom Line: The acetonitrile fractionated plasma proteins were subjected to reduction/alkylation and trypsin digestion followed by liquid chromatography and tandem mass spectrometry, which showed the absence of protocadherin 15, vascular endothelial growth factor-C, and certain transcription and ubiquitin-mediated proteolytic factors in FHS-prone birds.Using this experimental model, we analyzed the plasma peptides and proteins from normal and FHS-prone chickens by mass spectrometry to identify differentially expressed peptides and proteins.We found two peptides, both derived from apolipoprotein A-I, quantitatively elevated and two proteins, protocadherin 15 and VEGF-C, that were conspicuously absent in FHS-susceptible birds.

View Article: PubMed Central - PubMed

Affiliation: Cell & Molecular Biology Program and Poultry Science Department, University of Arkansas, Fayetteville, AR, USA.

ABSTRACT

Unlabelled: Femoral head separation (FHS) is an idiopathic bone problem that causes lameness and production losses in commercial poultry. In a model of prednisolone-induced susceptibility to FHS, the changes in plasma proteins and peptides were analyzed to find possible biomarkers. Plasma samples from control and FHS-susceptible birds were depleted of their high abundance proteins by acetonitrile precipitation and were then subjected to cation exchange and reverse-phase (RP) fractionations. Analysis with matrix assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) showed several differentially expressed peptides, two of which were isolated by RP-HPLC and identified as the fragments of apolipoprotein A-I. The acetonitrile fractionated plasma proteins were subjected to reduction/alkylation and trypsin digestion followed by liquid chromatography and tandem mass spectrometry, which showed the absence of protocadherin 15, vascular endothelial growth factor-C, and certain transcription and ubiquitin-mediated proteolytic factors in FHS-prone birds. It appears that prednisolone-induced dyslipidemia, vascular, and tissue adhesion problems may be consequential to FHS. Validity of these biomarkers in our model and the natural disease must be verified in future using traditional approaches.

Biomarker insights: Lameness because of femoral head separation (FHS) is a production and welfare problem in the poultry industry. Selection against FHS requires identification of the birds with subclinical disease with biomarkers from a source such as blood. Prednisolone can induce femoral head problems and predisposition to FHS. Using this experimental model, we analyzed the plasma peptides and proteins from normal and FHS-prone chickens by mass spectrometry to identify differentially expressed peptides and proteins. We found two peptides, both derived from apolipoprotein A-I, quantitatively elevated and two proteins, protocadherin 15 and VEGF-C, that were conspicuously absent in FHS-susceptible birds.

No MeSH data available.


Related in: MedlinePlus

(A) Protein sequence of chicken apolipoprotein A-I and the corresponding regions representing the 7304 Da (underlined) and the 3203 Da peptide (shown in bold) and (B) the possible cleavage sites that may generate these peptides, predicted by PROSPER online software.
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f4-bmi-10-2015-001: (A) Protein sequence of chicken apolipoprotein A-I and the corresponding regions representing the 7304 Da (underlined) and the 3203 Da peptide (shown in bold) and (B) the possible cleavage sites that may generate these peptides, predicted by PROSPER online software.

Mentions: The peptide profiles of CTRL and FHS samples obtained by RP and SCX fractionation methods are shown in Supplementary Tables S1 and S2. Although several peaks between the two groups were different as per CPT analysis, we isolated only two peptides m/z 7304 and m/z 3203 (Figs. 2A and 3A) by RP-HPLC both of which were the fragments of chicken apolipoprotein A-I (APOA1) derived from its C-terminal region. The peptide m/z 3203 was internal to m/z 7304 sequence as shown by MS and MS/MS results (Figs. 2B, 2C, 3B, and 3C). In silico analysis using PROSPER23 suggested a probability of the generation of these fragments by the action of cysteine and serine proteases, respectively (Figs. 4A and 4B).


Prednisolone-induced predisposition to femoral head separation and the accompanying plasma protein changes in chickens.

Packialakshmi B, Liyanage R, Lay J, Okimoto R, Rath N - Biomark Insights (2015)

(A) Protein sequence of chicken apolipoprotein A-I and the corresponding regions representing the 7304 Da (underlined) and the 3203 Da peptide (shown in bold) and (B) the possible cleavage sites that may generate these peptides, predicted by PROSPER online software.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4295844&req=5

f4-bmi-10-2015-001: (A) Protein sequence of chicken apolipoprotein A-I and the corresponding regions representing the 7304 Da (underlined) and the 3203 Da peptide (shown in bold) and (B) the possible cleavage sites that may generate these peptides, predicted by PROSPER online software.
Mentions: The peptide profiles of CTRL and FHS samples obtained by RP and SCX fractionation methods are shown in Supplementary Tables S1 and S2. Although several peaks between the two groups were different as per CPT analysis, we isolated only two peptides m/z 7304 and m/z 3203 (Figs. 2A and 3A) by RP-HPLC both of which were the fragments of chicken apolipoprotein A-I (APOA1) derived from its C-terminal region. The peptide m/z 3203 was internal to m/z 7304 sequence as shown by MS and MS/MS results (Figs. 2B, 2C, 3B, and 3C). In silico analysis using PROSPER23 suggested a probability of the generation of these fragments by the action of cysteine and serine proteases, respectively (Figs. 4A and 4B).

Bottom Line: The acetonitrile fractionated plasma proteins were subjected to reduction/alkylation and trypsin digestion followed by liquid chromatography and tandem mass spectrometry, which showed the absence of protocadherin 15, vascular endothelial growth factor-C, and certain transcription and ubiquitin-mediated proteolytic factors in FHS-prone birds.Using this experimental model, we analyzed the plasma peptides and proteins from normal and FHS-prone chickens by mass spectrometry to identify differentially expressed peptides and proteins.We found two peptides, both derived from apolipoprotein A-I, quantitatively elevated and two proteins, protocadherin 15 and VEGF-C, that were conspicuously absent in FHS-susceptible birds.

View Article: PubMed Central - PubMed

Affiliation: Cell & Molecular Biology Program and Poultry Science Department, University of Arkansas, Fayetteville, AR, USA.

ABSTRACT

Unlabelled: Femoral head separation (FHS) is an idiopathic bone problem that causes lameness and production losses in commercial poultry. In a model of prednisolone-induced susceptibility to FHS, the changes in plasma proteins and peptides were analyzed to find possible biomarkers. Plasma samples from control and FHS-susceptible birds were depleted of their high abundance proteins by acetonitrile precipitation and were then subjected to cation exchange and reverse-phase (RP) fractionations. Analysis with matrix assisted laser desorption ionization-time-of-flight mass spectrometry (MALDI-TOF-MS) showed several differentially expressed peptides, two of which were isolated by RP-HPLC and identified as the fragments of apolipoprotein A-I. The acetonitrile fractionated plasma proteins were subjected to reduction/alkylation and trypsin digestion followed by liquid chromatography and tandem mass spectrometry, which showed the absence of protocadherin 15, vascular endothelial growth factor-C, and certain transcription and ubiquitin-mediated proteolytic factors in FHS-prone birds. It appears that prednisolone-induced dyslipidemia, vascular, and tissue adhesion problems may be consequential to FHS. Validity of these biomarkers in our model and the natural disease must be verified in future using traditional approaches.

Biomarker insights: Lameness because of femoral head separation (FHS) is a production and welfare problem in the poultry industry. Selection against FHS requires identification of the birds with subclinical disease with biomarkers from a source such as blood. Prednisolone can induce femoral head problems and predisposition to FHS. Using this experimental model, we analyzed the plasma peptides and proteins from normal and FHS-prone chickens by mass spectrometry to identify differentially expressed peptides and proteins. We found two peptides, both derived from apolipoprotein A-I, quantitatively elevated and two proteins, protocadherin 15 and VEGF-C, that were conspicuously absent in FHS-susceptible birds.

No MeSH data available.


Related in: MedlinePlus