Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

The age-dependent changes in the number of3H-ouabain binding sites in mammalian skeletal muscle


The influence of age on the binding of3H-ouabain in skeletal muscle has been characterized in rats, mice and guinea pigs. Measurements performed using biopsies and intact fibers obtained from different types of rat muscles showed that from birth to the 4th week of life, the number of3H-ouabain binding sites per unit weight increases up to 5-fold, followed by almost the same relative decrease to a plateau around 250 pmol/g wet wt at an age of 22 weeks. These changes were not associated with any major alterations in apparentK D (1.7–3.1×10−7M) dissociation rate or heterogeneity in binding characteristics. Measurements of 3-O-methylfluorescein phosphatase activity, an enzyme activity which is closely correlated to the Na−K-ATPase activity, confirmed the3H-ouabain binding data.

In mice, the number of3H-ouabain binding sites showed similar, albeit less pronounced changes with age, a maximum being reached at the 4th week of life. In guinea pigs, the number of3H-ouabain binding sites per unit weight decreased by 60% from birth to maturity.

The results indicate that the early development and differentiation of individual skeletal muscles is associated with a marked increase in the number of Na−K-pumps (when expressed as pmol/muscle), until at maturity a plateau is reached. However, when expressed as pmol/g wet wt the increase is followed by a decrease to a plateau. This may in part account for the relatively low digitalis sensitivity seen in infants as compared to newborn and mature individuals.

This is a preview of subscription content, log in to check access.


  1. Aalkjær C, Kjeldsen K, Nørgaard Aa, Clausen T, Mulvany JM (1984)3H-ouabain binding sites and sodium content in resistance vessels and skeletal muscle of spontaneously hypertensive and potassium depleted rats. Hypertension (submitted)

  2. Asano Y, Liberman UA, Edelmann IS (1976) Relationship between Na+-dependent respiration and Na+−K+-adenosine triphosphatase activity in rat skeletal muscle. J Clin Invest 57:368–379

  3. Biron R, Burger A, Chinet A, Clausen T, Dubois-Ferriere R (1979) Thyroid hormones and the energetics of active sodium-potassium transport in mammalian skeletal muscles. J Physiol 27:47–60

  4. Clausen T, Hansen O (1974) Ouabain binding and Na+−K+ transport in rat muscle cells and adipocytes. Biochem Biophys Acta 345:387–404

  5. Clausen T, Hansen O (1977) Active Na−K transport and the rate of ouabain binding. The effect of insulin and other stimuli on skeletal muscle and adipocytes. J Physiol 270:415–430

  6. Clausen T, Sellin LC, Thesleff S (1981) Quantitative changes in ouabain binding after denervation and during reinnervation of mouse skeletal muscle. Acta Physiol Scand III:373–375

  7. Clausen T, Hansen O, Kjeldsen K, Nørgaard Aa (1982) Effect of age, potassium depletion and denervation on specific displaceable3H-ouabain binding in rat skeletal muscle in vivo. J Physiol 333:367–381

  8. Clausen T, Kjeldsen K, Nørgaard Aa (1983) Effecgs of denervation on sodium, potassium and3H-ouabain binding in muscles of normal and potassium-depleted rats. J Physiol 345:123–134

  9. Crettaz M, Prentki M, Zaninetti D, Jeanrenaud B (1980) Insulin resistance in soleus muscle from obese zucker rats. Biochem J 186:525–534

  10. DeFronzo RA, Bia M, Smith D (1982) Clinical disorders of hyperkalemia. Ann Rev Med 33:521–554

  11. Desaiah D, Mishra SK, Hobson M (1981) Quantitative measurements of3H-ouabain binding to human skeletal muscle cryostat sections. J Neurol Sci 51:457–464

  12. Desnuelle C, Lombet A, Serratrice G, Lazdunski M (1982) Sodium channel and sodium pump in normal and pathological muscles from patients with myotonic muscular dystrophy and lower motor neuron impairment. J Clin Invest 69:358–367

  13. De Villafranca GW (1954) Adenosinetriphosphatase activity in developing rat muscle. J Exp Zool 127:367–388

  14. Drachman DB, Johnston DM (1973) Development of a mammalian fast muscle: Dynamic and biochemical properties correlated. J Physiol 234:29–42

  15. Edge MB (1970) Development of apposed sarcoplasmic reticulum at the T system and sarcolemma and the change in orientation of triads in rat skeletal muscle. Dev Biol 23:634–650

  16. Erdman E, Philipp G, Scholtz H (1980) Cardiac gylcoside receptor, (Na+−K+)-ATPase activity and force of contraction in rat heart. Biochem Pharmacol 29:3219–3229

  17. Hansen O, Skou JC (1973) A study of the influence of the concentration of Mg2+, Pi, K+, Na+, and tris on (Mg2++Pi)-supported g-strophanthin binding to (Na++K+)-activated ATPase from ox brain. Biochim Biophys Acta 311:51–66

  18. Hnik P, Holas M, Krekule I, Kriz N, Mejsnar J, Smiesko V, Ujec E, Vyskocil F (1976) Work-induced potassium changes in skeletal muscle and effluent venous blood assessed by liquid ion-exchanger microelectrodes. Pflügers Arch 362:85–94

  19. Kjeldsen K, Nørgaard Aa, Clausen T (1982) Age-dependent changes in the number of3H-ouabain binding sites in rat soleus muscle. Biochim Biophys Acta 686:253–256

  20. Kjeldsen K, Nørgaard Aa, Clausen T (1984a) Effect of K-depletion on3H-ouabain binding and Na−K-contents in mammalian skeletal muscle. Acta Physiol Scand (in press)

  21. Kjeldsen K, Nørgaard Aa, Gøtzsche CO, Thomassen A, Clausen T (1984b) The effect of thyroid function on the number of Na−K-pumps in human skeletal muscle. Lancet 8393:8–10

  22. Kjeldsen K, Nørgaard Aa, Clausen T (1984c) Changes in the number of Na−K-pumps (digitalis-receptors) in skeletal muscle may in part account for the variations in digitalis toxicity with age, K-deficiency and the thyroid status. Eur Heart J 5 (Abstr Suppl 1), 43

  23. Neill CA (1965) The use of digitalis in infants and children. Prog. Cardiovasc Dis 7:399–416

  24. Nørgaard Aa, Kjeldsen K, Clausen T (1981) Potassium depletion decreases the number of3H-ouabain binding sites and the active Na−K-transport in skeletal muscle. Nature 293:739–741

  25. Nørgaard Aa, Kjeldsen K, Hansen O, Clausen T (1983) A simple and rapid method for the determination of the number of3H-ouabain binding sites in biopsies of skeletal muscle. Biochim Biophys Res Commun 111:319–325

  26. Nørgaard Aa, Kjeldsen K, Clausen T (1984a) A method for the determination of the total number of3H-ouabain binding sites in biopsies of human skeletal muscle. J Scand Clin Lab Invest (in press)

  27. Nørgaard Aa, Kjeldsen K, Hansen O (1984b) (Na++K+)-ATPase activity of crude homogenates of rat skeletal muscle as estimated from their K+-dependent 3-O-methylfluorescein phosphatase activity. Biochim Biophys Acta 770:203–209

  28. Sejersted OM, Medbø JI, Hermansen L (1982) Metabolic acidosis and changes in water and electrolyte balance after maximal exercise. In: Metabolic acidosis. Pitman Books Ltd. London. Ciba Foundation Symp 87:153–167

  29. Sperelakis N (1972) (Na+, K+)-ATPase activity of embryonic chick heart and skeletal muscles as a function of age. Biochim Biophys Acta 266:230–237

  30. Vigne P, Frelin C, Lazdunski M (1982) Ontogeny of the (Na+, K+)-ATPase during chick skeletal myogenesis. J Biol Chem 257:5380–5384

  31. Wettrell G, Andersson K-E (1977) Clinical pharmacokinetics of digoxin in infants. Clin Pharmacokinetics 2:17–31

Download references

Author information

Correspondence to Keld Kjeldsen.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Kjeldsen, K., Nørgaard, A. & Clausen, T. The age-dependent changes in the number of3H-ouabain binding sites in mammalian skeletal muscle. Pflugers Arch. 402, 100–108 (1984).

Download citation

Key words

  • 3H-Ouabain binding
  • Sodium-potassium pumps
  • Skeletal muscle
  • Age
  • Digitalis glycosides
  • 3-O-methylfluorescein phosphatase