Sequence analysis and expression profiling of the equine ACTN3 gene during exercise in Arabian horses

The ACTN3 gene codes for α-actinin-3, a protein localized in the Z-line in the skeletal muscle. Actinin-3 is critical in anchoring the myofibrillar actin filaments and plays a key role in muscle contraction. ACTN3 (α-actinin-3) cross-links glycogen phosphorylase (GP), which is the key enzyme catalysing glycogen metabolism. The aim of present study was to establish the expression level of the ACTN3 gene (for both isoforms separately and together in the gene expression analysis) in the gluteus medius muscle in order to verify if the α-actinin-3 gene can be related to training intensity in Arabian horses. A structural analysis of the ACTN3 gene was performed simultaneously to identify polymorphisms that can be related to racing performance traits. Our results showed the significant decrease (p < 0.05) of ACTN3 expression in the skeletal muscle of Arabian horses during the training periods preparing for flat-racing, and this decrease differed by the intensity of the exercises. The highest mRNA abundance measured for all ACTN3 genes was detected in the muscle of untrained horses, while the lowest expression was identified at the end of the racing season when horses had fully adapted to the physical effort. This gene expression profile was confirmed for both ACTN3 isoforms. The analysis of the ACTN3 sequence allowed us to identify 14 polymorphisms, which were localized in the promoter region, the 5'UTR (7 SNPs), exons (2 SNPs) and introns (5 SNPs). Two of them, a novel c.2334C>T - splice variant and the g.1104G>A polymorphism in the promoter region, were proposed as the causative mutations that might affect gene expression. The presented gene expression analyses indicated the significant role of the ACTN3 gene in adaptation to physiological effort in horses. Due to previous reports and our findings, further studies should be conducted to verify the usage of the ACTN3 gene as a potential genetic marker for determining exercise performance in Arabian horses and other horse breeds.