Prolonged submaximal exercise induces isoform-specific Na+-K+-ATPase mRNA and protein responses in human skeletal muscle

• Published in: American journal of physiology. Regulatory, integrative and comparative physiology integrative and comparative physiology, 2006-02, Vol.290 (2), p.R414-R424
• Main Authors: Murphy, K. T, Petersen, A. C, Goodman, C, Gong, X, Leppik, J. A, Garnham, A. P, Cameron-Smith, D, Snow, R. J, McKenna, M. J
• Format: Article
• Language: English
• Subjects: Adult; Exercise - physiology; Female; Gene Expression Regulation, Enzymologic; Humans; Male; Muscle Proteins - genetics; Muscle Proteins - metabolism; Muscle, Skeletal - enzymology; Muscle, Skeletal - metabolism; Ouabain - metabolism; Protein Binding; Protein Isoforms - genetics; Protein Isoforms - metabolism; Protein Subunits - metabolism; RNA, Messenger - genetics; RNA, Messenger - metabolism; Sodium-Potassium-Exchanging ATPase - genetics; Sodium-Potassium-Exchanging ATPase - metabolism
• ISSN: 0363-6119; 1522-1490
• DOI: 10.1152/ajpregu.00172.2005

1 Muscle, Ions, and Exercise Group, School of Human Movement, Recreation, and Performance, Centre for Aging, Rehabilitation, Exercise, and Sport Science (CARES), Victoria University of Technology, Melbourne; Australia; 2 Exercise, Muscle, and Metabolism Unit, School of Exercise and Nutrition Sciences, Deakin University, Melbourne; Australia Submitted 10 March 2005 ; accepted in final form 15 September 2005 This study investigated effects of prolonged submaximal exercise on Na + -K + -ATPase mRNA and protein expression, maximal activity, and content in human skeletal muscle. We also investigated the effects on mRNA expression of the transcription initiator gene, RNA polymerase II (RNAP II), and key genes involved in protein translation, eukaryotic initiation factor-4E (eIF-4E) and 4E-binding protein 1 (4E-BP1). Eleven subjects (6 men, 5 women) cycled at 75.5% (SD 4.8%) peak O 2 uptake and continued until fatigue. A vastus lateralis muscle biopsy was taken at rest, fatigue, and 3 and 24 h postexercise. We analyzed muscle for Na + -K + -ATPase 1 , 2 , 3 , 1 , 2 , and 3 , as well for RNAP II, eIF-4E, and 4E-BP1 mRNA expression by real-time RT-PCR and Na + -K + -ATPase isoform protein abundance using immunoblotting. Muscle homogenate maximal Na + -K + -ATPase activity was determined by 3 -O -methylfluorescein phosphatase activity and Na + -K + -ATPase content by [ 3 H]ouabain binding. Cycling to fatigue [54.5 (SD 20.6) min] immediately increased 3 ( P = 0.044) and 2 mRNA ( P = 0.042) by 2.2- and 1.9-fold, respectively, whereas 1 mRNA was elevated by 2.0-fold at 24 h postexercise ( P = 0.036). A significant time main effect was found for 3 protein abundance ( P = 0.046). Exercise transiently depressed maximal Na + -K + -ATPase activity ( P = 0.004), but Na + -K + -ATPase content was unaltered throughout recovery. Exercise immediately increased RNAP II mRNA by 2.6-fold ( P = 0.011) but had no effect on eIF-4E and 4E-BP1 mRNA. Thus a single bout of prolonged submaximal exercise induced isoform-specific Na + -K + -ATPase responses, increasing 1 , 3 , and 2 mRNA but only 3 protein expression. Exercise also increased mRNA expression of RNAP II, a gene initiating transcription, but not of eIF-4E and 4E-BP1, key genes initiating protein translation. gene expression; RNA polymerase II; 3 -O -methylfluorescein phosphatase; [ 3 H]ouabain binding Address for reprint requests and other correspondence: M. J. McKenna, School of Human Movement, Recreation and Performance (F022