Novel markers for tying-up in horses by proteomics analysis of equine muscle biopsies.
The aim of the study was to identify new biomarkers for acute tying-up in horses. Skeletal muscle biopsies were taken from 3 horses suffering from acute tying-up and 3 healthy horses. We performed 2D gel electrophoresis and mass spectrometry for identification of proteins that are differentially expressed in tying-up. 2D gel electrophoresis of skeletal muscle sequential extracts yielded more than 350 protein spots on each gel, of which 14 were differentially expressed more than two-fold (p<0.05). In-gel digestion followed by peptide mass fingerprinting enabled identification of three significantly increased proteins: alpha actin, tropomyosin alpha chain and creatine kinase M chain (CKM). CKM was represented by multiple spots probably due to posttranslational modification, one of which appeared to be unique for tying-up. Since changes in the rates of synthesis and degradation of proteins are likely to lead to pathological conditions, identification of differentially expressed proteins in acute tying-up might result in the finding of more specific diagnostic markers and in new hypotheses for the common mechanisms that result in this condition. Our findings point to a specific isoform of CKM as a novel biomarker for tying-up suggesting that altered energy distribution within muscle cells is part of the disease etiology.
Bouwman
Freek G
FG
Department of Human Biology, Maastricht University Medical Centre, The Netherlands.
van Ginneken
Mireille M E
MM
van der Kolk
Johannes H
JH
van Breda
Eric
E
Mariman
Edwin C M
EC
D000818:Animals; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D006734:Horse Diseases / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism; D006736:Horses / Q000235:genetics / Q000378:metabolism*; D009124:Muscle Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism / Q000473:pathology; D020914:Myopathies, Structural, Congenital / Q000175:diagnosis* / Q000235:genetics / Q000378:metabolism / Q000662:veterinary*; D040901:Proteomics / Q000379:methods*
D015415:Biomarkers; D009124:Muscle Proteins