Na+-K+-ATPase in rat skeletal muscle: content, isoform, and activity characteristics

• Published in: Journal of applied physiology (1985) Vol. 96; no. 1; pp. 316 - 326
• Main Authors: Fowles, J. R, Green, H. J, Ouyang, J
• Format: Journal Article
• Language: English
• Published: Bethesda, MD Am Physiological Soc 01.01.2004; American Physiological Society
• Subjects: Animals; Biological and medical sciences; Cell Membrane - enzymology; Enzyme Activation - physiology; Enzyme Inhibitors - metabolism; Enzyme Inhibitors - pharmacology; Enzymes; Fundamental and applied biological sciences. Psychology; Isoenzymes - metabolism; Male; Muscle, Skeletal - enzymology; Muscular system; Organ Specificity; Ouabain - metabolism; Ouabain - pharmacology; Rats; Rats, Wistar; Rodents; Sodium; Sodium-Potassium-Exchanging ATPase - metabolism; Tritium; fiber types; Molecular form; Rat; Enzyme; Cyclism; Rodentia; isoforms; K; Striated muscle; cation cycling; Sport; Vertebrata; Mammalia; Na; sodium pump; Enzymatic activity; enzyme activity; Hydrolases; exchanging ATPase; Cations
• ISSN: 8750-7587; 1522-1601
• DOI: 10.1152/japplphysiol.00745.2002

Department of Kinesiology, University of Waterloo, Waterloo, Ontario, Canada N2L 3G1 Submitted 12 August 2002 ; accepted in final form 9 July 2003 The purpose of this study was to investigate the hypothesis that muscle Na + -K + -ATPase activity is directly related to Na + -K + -ATPase content and the content of the 2 -catalytic isoform in muscles of different fiber-type composition. To investigate this hypothesis, tissue was sampled from soleus (Sol), red gastrocnemius (RG), white gastrocnemius (WG), and extensor digitorum longus (EDL) muscles at rest from 38 male Wistar rats weighing 413 ± 6.0 g (mean ± SE). Na + -K + -ATPase activity was determined in homogenates (Hom) and isolated crude membranes (CM) by the regenerating ouabain-inhibitable hydrolytic activity assay (ATPase) and the 3- O -methylfluorescein K + -stimulated phosphatase (3- O -MFPase) assay in vitro. In addition, Na + -K + -ATPase content (B max ) and the distribution of 1 -, 2 -, 1 -, and 2 -isoforms were determined by [ 3 H]ouabain binding and Western blot, respectively. For the ATPase assay, differences ( P < 0.05) in enzyme activity between muscles were observed in Hom (EDL > WG) and in CM (Sol > EDL = WG). For the 3- O -MFPase assay, differences ( P < 0.05) were also found for Hom (Sol > RG = EDL > WG) and CM (Sol = WG > RG). For B max , differences in the order of RG = EDL > Sol = WG ( P < 0.05) were observed. Isoform distribution was similar between Hom and CM and indicated in CM, a greater density ( P < 0.05) of 1 in Sol than WG and EDL ( P < 0.05), but more equal distribution of 2 between muscles. The 1 was greater ( P < 0.05) in Sol and RG, and the 2 was greater in EDL and WG ( P < 0.05). Over all muscles, the correlation ( r ) between Hom 3- O -MFPase and B max was 0.45 ( P < 0.05) and between Hom 2 and B max , 0.59 ( P < 0.05). The 1 distribution correlated to Hom 3- O -MFPase ( r = 0.79, P < 0.05) CM ATPase ( r = 0.69, P < 0.005) and CM 3- O -MFPase activity ( r = 0.32, P < 0.05). The 2 distribution was not correlated with any of the Na + -K + -ATPase activity measurements. The results indicate generally poor relationships between activity and total pump content and 2 isoform content of the Na + -K + -ATPase. Several factors, including the type of preparation and the type of assay, appear important in this regard. sodium pump; fiber types; cation cycling; enzyme activity; isoforms Address for reprint requests and other correspondence: H. J. Green, Dept. of Kinesiology, Univ. of Waterloo, Waterloo, ON, Canada N2L 3G1 (E-mail: green{at}healthy.uwaterloo.ca ).