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The effect of temperature on the pH stability of myosin ATPase as demonstrated histochemically

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Summary

The histochemical reaction for actomyosin adenosine triphosphatase (ATPase) and its lability to pH variations characterize two major categories of fibre types in mixed mammalian skeletal muscles. These are designated as the “Type I” and “Type II” fibres. Myosin ATPase of Type I fibres has been shown to be relatively acid stable but alkali labile while that of Type II fibres is relatively alkali stable but acid labile. In this study we have investigated the effect of preincubation at varying pH and temperatures on the histochemical demonstration of myosin ATPase in animal and human mixed skeletal muscles. Sections fixed in cold fixative at either high (10.4) or low (4.35) pH showed identical staining patterns. When alternate serial sections were preincubated in either cold (4°C) acidic or cold alkaline solutions, only the Type II fibres were intensely stained. After alkaline preincubation at room temperature (19–21°C) varying degrees of ATPase inhibition were observed in Type II fibres while Type I fibres remained unstained. Acidic preincubation at room temperature (19–20°C) resulted in the enhancement of staining of the Type I fibres but variable inhibition of activity was found in the Type II fibres. This study demonstrates that during preincubation, pH is not the only critical factor and the specific temperature of the acidic or alkaline preincubation bath is a major contributing factor in the histochemical demonstration of myosin ATPase. It also demonstrates that a temperature dependent loss of myosin ATPase activity in the frozen sections of skeletal muscle following acidic or alkaline pretreatment is consistent with previous biochemical reports on purified actomyosin (Guth and Samaha, 1969). Moreover, the heterogeneity can be displayed within both Type II white and Type II red muscle fibres when examined for myosin ATPase after acid preincubation.

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References

  • Aronson, J. F.: A melting point for the birefringent component of muscle. J. Cell Biol. 30, 453–646 (1966)

    Google Scholar 

  • Ashmore, C. R., Doerr, L.: Comparative aspects of muscle fiber types in different species. Exp. Neurol. 31, 408–418 (1971)

    Google Scholar 

  • Ashmore, C. R., Tompkins, G., Doerr, L.: Postnatal development of muscle fiber types in domestic animals. J. Anim. Sci. 34, 37–41 (1972)

    Google Scholar 

  • Barka, T., Anderson, P.: Histochemistry, theory, practice and bibliography, p. 313. New York: Harper Row Publishers, Inc. 1963

    Google Scholar 

  • Bendall, J. R.: The myofibrillar ATPase activity of various animals in relation to ionic strength and temperature. In: Biochemistry of muscle contraction, Gergely, J. (ed.), p. 448–451. Boston: Little Brown and Co. 1964

    Google Scholar 

  • Brahms, J., Brenzer, J.: Interaction of myosin A with ions. Arch. Biochem. Biophys. 95, 219–228 (1961)

    Google Scholar 

  • Brooke, M. H., Kaiser, K. K.: Some comments on the histochemical characterization of muscle adenosine triphosphatase. J. Histochem. Cytochem. 17, 431–432 (1969)

    Google Scholar 

  • Brooke, M. H., Kaiser, K. K.: Three “myosin adenosine triphosphatase” systems: The nature of their pH lability and sulphydryl dependence. J. Histochem. Cytochem. 18, 670–672 (1970a)

    Google Scholar 

  • Brooke, M. H., Kaiser, K. K.: Muscle fibre types: How many and what kind? Arch. Neurol. (Chic.) 23, 369–379 (1970b)

    Google Scholar 

  • Brooke, M. H., Williamson, E., Kaiser, K. K.: The behaviour of four fiber types in developing and reinnervated muscle. Arch. Neurol. (Chic.) 25, 360–366 (1971)

    Google Scholar 

  • Burke, R. E., Levine, D. N., Zajac, F. E., Tsairis, P., Engel, W. K.: Mammalian motor units: Physiological-histochemical correlation in three types in cat gastrocnemius. Science 174, 709–712 (1971)

    Google Scholar 

  • Close, R.: Properties of motor units in fast and slow skeletal muscles of the rat. J. physiol. (Lond.) 193, 45–55 (1967)

    Google Scholar 

  • Davies, A. S.: Postnatal changes in the histochemical fibre types of porcine skeletal muscle. J. Anat. (Lond.) 113, 213–240 (1972)

    Google Scholar 

  • Drews, G. A., Engel, W. K.: Reversal of the ATPase reaction in muscle fibers by EDTA. Nature (Lond.) 212, 1551–1553 (1966)

    Google Scholar 

  • Edgerton, V. R., Gerchman, L., Carrow, R.: Histochemical changes in rat skeletal muscle after exercise. Exp. Neurol. 24, 110–123 (1969)

    Google Scholar 

  • Edgerton, V. R., Simpson, D. R.: The intermediate muscle fiber of rats and guinea pigs. J. Histochem. Cytochem. 17, 828–838 (1969)

    Google Scholar 

  • Edgerton, V. R., Simpson, D. R.: Dynamic and metabolic relationship in the rat extensor digitorum longus muscle. Exp. Neurol. 30, 374–376 (1971)

    Google Scholar 

  • Engel, W. K.: Selective and non-selective susceptibility of muscle fiber types. Arch. Neurol. (Chic.) 22, 97–117 (1970)

    Google Scholar 

  • Gauthier, G. F.: On the relationship of ultrastructural and cytochemical features to color in mammalian skeletal muscle. Z. Zellforsch. 95, 462–482 (1969)

    Google Scholar 

  • Gauthier, G. F., Padykula, H. A.: Cytological studies of fiber types in skeletal muscle. A comparative study of the mammalian diaphragm. J. Cell Biol. 28, 333–354 (1966)

    Google Scholar 

  • Gontefongea, R.: Influence of pH and temperature on solubility of muscle protein in pigs. Ann. Biol. anim. 11, 233–244 (1971)

    Google Scholar 

  • Guth, L., Samaha, F. J.: Qualitative differences between actomyosin ATPase of slow and fast mammalian muscle. Exp. Neurol. 25, 138–152 (1969)

    Google Scholar 

  • Guth, L., Samaha, F. J.: Procedure for the histochemical demonstration of actomyosin ATPase. Exp. Neurol. 28, 365–367 (1970)

    Google Scholar 

  • Guth, L., Samaha, F. J., Albers, R. W.: The neural regulation of some pheotypic differences between the fiber types of mammalian skeletal muscle. Exp. Neurol. 26, 126–135 (1970)

    Google Scholar 

  • Guth, L., Yellin, H.: The dynamic nature of the so-called “Fiber types” of mammalian skeletal muscle. Exp. Neurol. 31, 277–300 (1971)

    Google Scholar 

  • Hoh, J. F. Y.: Electrophoresis of intact myosin in polyacrylamide gel. Proc. Aust. Biochem. Soc. 5, 20 (1972)

    Google Scholar 

  • Karpati, G., Engel, W. K.: Histochemical investigation of fiber type ratios with the myofibrillar ATPase reaction in normal and denervated skeletal muscles in guinea pig. Amer. J. Anat. 122, 145–156 (1968)

    Google Scholar 

  • Kawakami, H., Marita, J., Takahashi, K., Yasui, T.: Thermal denaturation of myosin, heavy mero-myosin and sub-fragment I. J. Biochem. 70, 635–648 (1971)

    Google Scholar 

  • Khan, M. A., Papadimitrion, J. M., Holt, P. G., Kakulas, B. A.: A calcium-citro-phosphate technique for the histochemical localization of myosin ATPase. Stain Technol. 47, 277–281 (1972a)

    Google Scholar 

  • Khan, M. A., Papadimitriou, J. M., Holt, P. G., Kakulas, B. A.: A modified histochemical technique for sarcoplasmic reticular ATPase. Histochemie 30, 329–333 (1972b)

    Google Scholar 

  • Khan, M. A., Papadimitriou, J. M., Holt, P. G., Kakulas, B. A.: A histochemical analysis of mammalian oxidative skeletal muscle fibers using the enzymes of energetic metabolism. Histochemie 33, 310–312 (1973a)

    Google Scholar 

  • Khan, M. A., Papadimitriou, J. M., Holt, P. G., Kakulas, B. A.: Further histochemical properties of rabbit skeletal muscle fibres. Histochemie 36, 173–183 (1973b)

    Google Scholar 

  • Lewis, M., Saraff, H. A.: The binding of ions to the muscle proteins. Measurements on the binding of potassium and sodium ions to myosin A, myosin B and actin. J. Amer. chem. Soc. 79, 2112–2117 (1957)

    Google Scholar 

  • McFarland, B. H., Inesi, G.: Solubilization of sarcoplasmic reticulum with Triton X-100. Arch. Biochem. Biophys. 145, 456–464 (1971)

    Google Scholar 

  • Okitani, A., Takogi, O., Fujimaki, M.: The changes of myosin B during storage of rabbit muscle. Part 4. Effect of temperature, pH and ionic strength on denaturation of myosin B solution. Agr. Biol. Chem. (Japan) 31, 939–944 (1967)

    Google Scholar 

  • Ouellet, L., Laidler, K. J., Morales, M. F.: Molecular kinetics of muscle adenosine triphosphatase. Arch. Biochem. Biophys. 39, 37–50 (1952)

    Google Scholar 

  • Padykula, H. A., Herman, E.: Factors affecting the activity of adenosine triphosphatase and other phosphatases as measured by histochemical techniques. J. Histochem. Cytochem. 3, 161–169 (1955)

    Google Scholar 

  • Penn, A. S., Cloak, R. A., Rowland, L. P.: Myosin from normal and dystrophic human muscle. Immunochemical and electrophoretic study. Arch. Neurol. (Chic.). 27, 159–173 (1972)

    Google Scholar 

  • Peter, J. B., Barnard, R. J., Edgerton, V. R., Gillespie, C. A., Stempel, K. E.: Metabolic profiles of three fibres types of skeletal muscle in guinea pigs and rabbits. Biochemistry 11, 2627–2633 (1972)

    Google Scholar 

  • Samaha, F. J.: Differences between the skeletal and heart types of actomyosin ATPase. Exp. Neurol. 35, 30–38 (1972)

    Google Scholar 

  • Samaha, F. J., Guth, L., Albers, R. W.: Differences between slow and fast muscle myosin. Adenosine triphosphatase activity and release of associated proteins by p-Chloromercuriphenylsulfonate. J. biol. Chem. 245, 219–244 (1970)

    Google Scholar 

  • Schiaffino, S., Bormioli, S. P.: Histochemical characterization of adenosine triphosphatases in skeletal muscle fibres by selective extraction procedures. J. Histochem. Cytochem. 21, 142–145 (1973)

    Google Scholar 

  • Stein, J. M., Padykula, H. A.: Histochemical classification of individual skeletal muscle fibers of the rat. Amer. J. Anat. 110, 103–123 (1962)

    Google Scholar 

  • Takahashi, K., Yasui, T., Hashimoto, Y., Tonomura, Y.: Physicochemical studies on denaturation of myosin adenosinetriphosphatase II. Changes in chromatographic profile and optical rotation. Arch. Biochem. Biophys. 99, 45–51 (1962)

    Google Scholar 

  • Yasui, T., Fukazawa, T., Hashimoto, Y., Kitigawa, S., Sasaki, A. T.: Denaturation of myosin B. J. Biochem. 45, 717–724 (1958)

    Google Scholar 

  • Yasui, T., Hashimoto, Y., Tonomura, Y.: Physicochemical studies on denaturation of myosinadenosine-triphosphatase. Arch. Biochem. Biophys. 87, 56–62 (1960)

    Google Scholar 

  • Yasui, T., Marita, J., Takahashi, K.: Inhibition by some neutral organic solutes of the denaturation of myosin A adenosine triphosphatase. J. Biochem. 60, 303–316 (1966)

    Google Scholar 

  • Yellin, H.: Differences in histochemical attributes between diaphragm and hindleg muscles of the rat. Anat. Rec. 173, 333–339 (1972)

    Google Scholar 

  • Yellin, H., Guth, L.: The histochemical classification of muscle fibers. Exp. Neurol. 26, 424–432 (1970)

    Google Scholar 

Download references

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, University of Western Australia, Perth, Western Australia

    M. A. Khan, J. M. Papadimitriou & B. A. Kakulas

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  1. M. A. Khan
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  2. J. M. Papadimitriou
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  3. B. A. Kakulas
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Khan, M.A., Papadimitriou, J.M. & Kakulas, B.A. The effect of temperature on the pH stability of myosin ATPase as demonstrated histochemically. Histochemistry 38, 181–194 (1974). https://doi.org/10.1007/BF00499665

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  • DOI: https://doi.org/10.1007/BF00499665

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