Packialakshmi B, Liyanage R, Lay Jo, Okimoto R, Rath Nc
Cell & Molecular Biology Program and Poultry Science Department, University of Arkansas, Fayetteville, AR, USA.
State wide Mass Spectrometry Facility, University of Arkansas, Fayetteville, AR, USA.
Biomark Insights. 2015 Jan 14;10:1-8. doi: 10.4137/BMI.S20268. eCollection 2015.
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.
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.
股骨头分离(FHS)是一种特发性骨骼问题,会导致商品家禽跛行和生产损失。在泼尼松龙诱导的FHS易感性模型中,分析血浆蛋白质和肽的变化以寻找可能的生物标志物。通过乙腈沉淀去除对照和FHS易感鸡的血浆样品中的高丰度蛋白质,然后进行阳离子交换和反相(RP)分级分离。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)分析显示了几种差异表达的肽,其中两种通过RP-HPLC分离并鉴定为载脂蛋白A-I的片段。对乙腈分级分离的血浆蛋白质进行还原/烷基化和胰蛋白酶消化,然后进行液相色谱和串联质谱分析,结果显示FHS易感鸡中不存在原钙黏蛋白15、血管内皮生长因子-C以及某些转录和泛素介导的蛋白水解因子。似乎泼尼松龙诱导的血脂异常、血管和组织黏附问题可能是FHS的后果。这些生物标志物在我们的模型和自然疾病中的有效性未来必须使用传统方法进行验证。
由于股骨头分离(FHS)导致的跛行是家禽业中的生产和福利问题。针对FHS进行选择需要从血液等来源使用生物标志物识别亚临床疾病的鸡。泼尼松龙可诱发股骨头问题和FHS易感性。利用这个实验模型,我们通过质谱分析了正常和FHS易感鸡的血浆肽和蛋白质,以鉴定差异表达的肽和蛋白质。我们发现两种均源自载脂蛋白A-I的肽定量升高,以及两种蛋白质,即原钙黏蛋白15和VEGF-C,在FHS易感鸡中明显缺失。