Proteomics technology used for discovery in recurrent airway obstruction in horses
Abstract
In order to evaluate the protein profile in the lungs of horses affected with recurrent airway obstruction (RAO), RAO-affected and control horses were subjected to an experimental exposure trial until the RAO-affected horses showed clinical signs of disease. Proteomic analysis was performed on bronchoalveolar lavage fluid (BALF) samples from both groups of horses and showed differential expression of 100 peaks. With a combination of LC-MS and LC-MS/MS techniques, identifications were made for 250 unique proteins from the BALF of RAO-affected and control horses. Transferrin and secretoglobin were chosen for validation with Western blot. Western blot was performed on BALF from the RAO-affected and control horses used in the experimental exposure trial and BALF from client-owned RAO-affected horses with naturally-induced clinical signs of disease and control horses from the same environments. Proteomics indicated that secretoglobin was not differentially expressed between the experimental exposure trial groups as confirmed by Western blot analysis. Validation with Western blot showed that client-owned RAO-affected horses had lower secretoglobin expression than client-owned control horses and control horses before experimental exposure. According to the proteomics data, transferrin was over-expressed in control horses after the experimental exposure compared to RAO-affected horses. While this comparison showed no statistical difference based on Western blot, transferrin was significantly over-expressed in control horses before experimental exposure compared to client-owned RAO-affected horses. In addition, both secretoglobin and transferrin band densities on Western blot were negatively correlated with airway obstruction and neutrophilic pulmonary inflammation. This study demonstrates that proteomics techniques can be used in the investigation of differentially-expressed proteins in the BALF of control horses and RAO-affected horses. The proteins identified as differentially expressed in this study including, but not limited to, secretoglobin and transferrin should undergo further evaluation for their use as biomarkers of disease, and potential targets of new therapeutic agents for RAO.
Degree
Ph.D.
Advisors
Couetil, Purdue University.
Subject Area
Veterinary services
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