Effects of acute exercise and long-term exercise on total Na+,K+-ATPase content and Na+,K+-ATPase isoform expression profile in equine muscle

Objective-To investigate the effects of acute exercise and long-term training on Na+,K+-ATPase content, mRNA isoforms, and protein concentration in equine muscle. Animals-6 Standardbreds. Procedures-Horses performed a bout of exercise on a treadmill before and after 18 weeks of combined interval and endurance training. Muscle biopsy specimens were obtained from vastus lateralis muscle (VLM) and pectoralis descendens muscle (PDM) before and after exercise. The Na+,K+-ATPase content, mRNA isoforms, and protein concentrations were determined by use of [H-3]ouabain binding, real-time PCR assay, and western blotting, respectively. Results-6 Na+,K+-ATPase mRNA isoforms were present in equine muscle, but only alpha 2 and beta 1 proteins were detected. Exercise before training resulted in increases of mRNA isoforms alpha 1, alpha 2, alpha 3, and beta 2 in VLM and alpha 1 and beta 3 in PDM. Training increased resting values for mRNA isoforms alpha 3 and beta 1 in VLM and beta 3 in PDM. The Na+,K+-ATPase, [3H]ouabain binding, and proteins of mRNA alpha 2 and beta 1 increased in VLM, whereas in PDM, only alpha 2 protein increased as a result of training. After training, effects of strenuous exercise on mRNA expression were no longer detectable. Conclusions and Clinical Relevance-Equine muscle contained all Na+,K+-ATPase mRNA isoforms, but only alpha 2 and beta 1 proteins could be detected. Expression of these isoforms changed as a result of strenuous exercise and long-term training, representing an adaptive response. Determination of Na+,K+-ATPase gene expression may be relevant for understanding alterations in excitability during neuromuscular diseases. (Am J Vet Res 2009-170:895-901)