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J Appl Physiol 87: 1132-1140, 1999;
8750-7587/99 $5.00
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Vol. 87, Issue 3, 1132-1140, September 1999

Age-associated differential expression of Na+-K+-ATPase subunit isoforms in skeletal muscles of F-344/BN rats

Xiwu Sun1, Murali Nagarajan1, Philip W. Beesley2, and Yuk-Chow Ng1

1 Department of Pharmacology, The Milton S. Hershey Medical Center, College of Medicine, The Pennsylvania State University, Hershey, Pennsylvania 17033-0850; and 2 Division of Biochemistry, School of Biological Sciences, Royal Holloway University of London, Egham, Surrey TW20 0EX, United Kingdom

Skeletal muscle expresses multiple isoforms of the Na+-K+-ATPase. Their expression has been shown to be differentially regulated under pathophysiological conditions. In addition, previous studies suggest possible age-dependent alterations in Na+-K+ pump function. The present study tests the hypothesis that advancing age is associated with altered Na+-K+-ATPase enzyme activity and isoform-specific changes in expression of the enzyme subunits. Red and white gastrocnemius (Gast) as well as soleus muscles of male Fischer 344/Brown Norway (F-344/BN) rats at 6, 18, and 30 mo of age were examined. Na+-K+-ATPase activity, measured by K+-stimulated 3-O-methylfluorescein phosphatase activity, increased by ~50% in a mixed Gast homogenate from 30-mo-old compared with 6- and 18-mo-old rats. Advancing age was associated with markedly increased alpha 1- and beta 1-subunit, and decreased alpha 2- and beta 2-subunit in red and white Gast. In soleus, there were similar changes in expression of alpha 1- and alpha 2-subunits, but levels of beta 1-subunit were unchanged. Functional Na+-K+-ATPase units, measured by [3H]ouabain binding, undergo muscle-type specific changes. In red Gast, high-affinity ouabain-binding sites, which are a measure of alpha 2-isozyme, increased in 30-mo-old rats despite decreased levels of alpha 2-subunit. In white Gast, by contrast, decreased levels of alpha 2-subunit were accompanied by decreased high-affinity ouabain-binding sites. Finally, patterns of expression of the four myosin heavy chain (MHC) isoforms (type I, IIA, IIX, and IIB) in these muscles were similar in the three age groups examined. We conclude that, in the skeletal muscles of F-344/BN rats, advancing age is associated with muscle type-specific alterations in Na+-K+-ATPase activity and patterns of expression of alpha - and beta -subunit isoforms. These changes apparently occurred without obvious shift in muscle fiber types, since expression of MHC isoforms remained unchanged. Some of the alterations occurred between middle-age (18 mo) and senescence (30 mo), and, therefore, may be attributed to aging of skeletal muscle.

aging; immunoblotting; ouabain binding; isozyme; sodium-potassium pump


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