Histochemistry

, Volume 73, Issue 4, pp 577–588 | Cite as

Alternative method for quantitative enzyme histochemistry of muscle fibers

Application of photographic densitometry combined with atomic absorption spectrophotometry
  • D. W. Sickles
  • R. E. McLendon
  • Th. H. Rosenquist
Article

Summary

The present study examines the use of photographic densitometry combined with atomic absorption spectrophotometry for the quantitation of enzyme activities (SDH and ATPase) in fresh frozen sections of rat tibialis anterior muscles. The technique eliminates some difficulties which are inherent in other methods. The reliability of the technique was found to be in the 98% range; the results were precise for all samples studied. The use of SDH to separate muscle fibers into “types” was found to be totally inaccurate since a full spectrum of activities was observed. ATPase activities could separate easily into two groups, but a continuum of ATPase activities was observed in the fast-twitch fibers. The simultaneous use of both enzymes was capable of separating the FG, FOG and SO fibers; however, variation within a single type was considerable and a great deal of information was lost when using any classification system. The continuum of SDH activities indicates the motor units are arranged as a spectrum of fatigue-resistant contractile units. The range of ATPase activities observed is comparable to ranges of motor unit contraction times emphasizing the importance of this enzyme in the regulation of contraction speed.

Keywords

Muscle Fiber ATPase Activity Motor Unit Atomic Absorption Spectrophotometry Tibialis Anterior Muscle 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barány M (1967) ATPase activity of myosin correlated with speed of muscle shortening. J Gen Physiol 50:197–218Google Scholar
  2. Burke RE (1978) Motor units: physiological: histochemical profiles, neural connectivity and functional specialization. Am Zool 18:127–134Google Scholar
  3. Burke RE, Tsairis P (1974) The correlation of physiological properties with histochemical characteristics in single muscle units. Ann NY Acad Sci 228:145–159Google Scholar
  4. Close RI (1972) Dynamic properties of mammalian skeletal muscles. Physiol Rev 52:129–197Google Scholar
  5. Edgerton VR (1976) Neuromuscular adaptation to power and endurance work. Can J Appl Sport Sci 1:49–58Google Scholar
  6. Eisenberg BR, Kuda AM (1976) Discrimination between fibre populations in mammalian skeletal muscle by using ultrastructural parameters. J Ultrastruct Res 54:76–88Google Scholar
  7. Essen B, Jansson E, Henriksson J, Taylor AW, Saltin B (1975) Metabolic characteristics of fibre types in human skeletal muscle. Acta Physiol Scand 95:153–165Google Scholar
  8. Frederick EC, Hamant MF, Rasmussen SA, Chan AK, Gaslow GE (1978) Correlation of histochemical and physiological properties of muscle units in striped skunk. Experientia 34:372–374Google Scholar
  9. Guth L, Samaha FJ (1970) Procedure for the histochemical demonstration of actomyosin ATPase. Exp Neurol 28:365–367Google Scholar
  10. Halkjaer-Kristensen J, Ingemann-Hansen T (1979) Microphotometric analysis of NADH-tetrazolium reductase and α-glycerophosphate dehydrogenase in human quadriceps muscle. Histochem J 11:127–136Google Scholar
  11. Hintz CS, Lowry CV, Kaiser KK, McKee D, Lowry OH (1980) Enzyme levels in individual rat muscle fibers. Am J Physiol 239:C58-C65Google Scholar
  12. Khan MA (1976) Histochemical characteristics of vertebrate striated muscle: a review. Prog Histochem Cytochem 8:1–48Google Scholar
  13. Lowry CV, Kimmey JS, Felder S, Chi MM-Y, Kaiser KK, Passoneau PN, Kirk KA, Lowry OH (1978) Enzyme patterns in single human muscle fibers. J Biol Chem 253:8269–8277Google Scholar
  14. Nachlas MM, Tsou K, Desouza E, Cheng C, Seligman AM (1957) Cytochemical demonstration of succinic dehydrogenase by the use of a new p-nitrophenyl substituted ditetrazole. J Histochem Cytochem 5:420–436Google Scholar
  15. Nolte J, Pette D (1972) Microphotometric determination of enzyme activity in single cells in cryostat sections. II. Succinate dehydrogenase lactate dehydrogenase and triosephosphate dehydrogenase activities in red, intermediate and white fibers of soleus and rectus femoris muscles of rat. J Histochem Cytochem 20:577–582Google Scholar
  16. Peter JB, Barnard RJ, Edgerton VR, Gillespie CA, Stempel KE (1972) Metabolic profiles of three fibre types of skeletal muscle in guinea pigs and rabbits. Biochemistry 11:2627–2633Google Scholar
  17. Pette D, Müller W, Leisner E, Vrbová G (1976) Time dependent effects on contractile properties, fibre population, myosin light chains and enzymes of energy metabolism in intermittently and continuously stimulated fast twitch muscles of the rabbit. Pflügers Arch 364:103–112Google Scholar
  18. Pool CW, Diegenbach PC, Scholten G (1979) Quantitative succinate dehydrogenase histochemistry I. a methodological study on a mammalian and fish muscle. Histochemistry 64:251–262Google Scholar
  19. Seidel JC (1967) Studies on myosin from red and white skeletal muscles of the rabbit. J Biol Chem 242:5623–5629Google Scholar
  20. Sickles DW, Pinkstaff CA (1981) Comparative histochemical study of prosimian primate hindlimb muscles. I. Muscle fiber types. Am J Anat 160:175–186Google Scholar
  21. Spamer C, Pette D (1977) Activity patterns of phosphofructokinase, glyceraldehydephosphate dehydrogenase, lactate dehydrogenase and malate dehydrogenase in microdissected fast and slow fibres from rabbit psoas and soleus muscle. Histochemistry 52:201–216Google Scholar
  22. Spamer C, Pette D (1979) Activities of malate dehydrogenase, 3-hydroxyacyl-CoA dehydrogenase and fructose-1,6 diphosphatase with regard to metabolic subpopulations of fast- and slow-twitch fibres in rabbit muscles. Histochemistry 60:9–21Google Scholar
  23. Spamer C, Pette D (1980) Metabolic subpopulations of rabbit skeletal muscle fibres. In: Pette D (ed) Plasticity of muscle. Walter de Gruyter, New York Berlin, pp 19–30Google Scholar
  24. Spurway NC (1980) Histochemical typing of muscle fibres by microphotometry. In: Pette D (ed) Plasticity of muscle. Walter de Gruyter, New York Berlin, pp 31–44Google Scholar
  25. Swatland HJ (1977) Transitional stages in the histochemical development of muscle fibres during post-natal growth. Histochem J 9:751–757Google Scholar
  26. Swatland HJ (1978) Comparison of red and white muscles by cytophotometry of their muscle fibre populations. Histochem J 10:349–360Google Scholar
  27. Taylor AW, Essen B, Saltin B (1974) Myosin ATPase in skeletal muscle of healthy men. Acta Physiol Scand 91:568–570Google Scholar
  28. Troyer H, Rosenquist TH (1975) Atomic absorption spectrophotometry applied to photographic densitometry. J Histochem Cytochem 23:941–944Google Scholar
  29. Van De Graaff KM, Frederick EC, Williamson RG, Goslow GE Jr (1977) Motor units and fiber types of primary ankle extensors of the skunk (Mephitis mephitis). J Neurophysiol 40:1424–31Google Scholar

Copyright information

© Springer-Verlag 1982

Authors and Affiliations

  • D. W. Sickles
    • 1
  • R. E. McLendon
    • 1
  • Th. H. Rosenquist
    • 1
  1. 1.Department of AnatomyMedical College of GeorgiaAugustaUSA

Personalised recommendations