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On the specificity of the histochemical technique for sarcoplasmic reticular adenosine triphosphatase: A light and electron microscopic study

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Summary

The specificity of the histochemical localization of the calcium activated adenosine triphosphatase (ATPase) activity of the sarcoplasmic reticulum (SR) at pH 7.4 was studied using a calcium-citro-phosphate technique. The latter involves the splitting of ATP by ATPase producing phosphate ions which then react with calcium and citrate to form an insoluble reaction product. This reaction product was detected by both light and electron microscopy. Light microscopic examination showed a darkly stained continuous reticular pattern of reaction product which surrounded individual myofibrils. This reticular pattern of reaction product was distinctly dissimilar to that found when the histochemical reactions for mitochondrial or myofibrillar ATPase were performed. Ultrastructural investigations demonstrated the presence of discrete foci of electron dense reaction product in close association with the membranes of the SR in striated muscle fibres. Only occasional flecks were seen in the vicinity of mitochondria or myofilaments. The possibility is considered that the reticular pattern of staining achieved by the calcium-citro-phosphate technique may reflect the distribution of the “extra ATPase” of the SR, an enzyme implicated in the process of calcium uptake and muscle relaxation.

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References

  • Andersson-Cedergren, E.: Ultrastructure of motor end plate and sarcoplasmic components of mouse skeletal muscle fiber as revealed by three-dimensional reconstruction from rapid serial sections. J. Ultrastruct. Res. 1, Suppl. (1959)

  • Barka, T., Anderson, P.: Histochemistry-theory, practice and bibliography, p. 313. New York: Hoeber Medical Division, Harper and Row Publ. Inc. 1963

    Google Scholar 

  • Barker, L. M., McPhillips, J. J., Lawrence, G. D., Doty, S. B., Pallante, S. L., Bills, C. E., Scott, W. W., Jr., Howard, J. E.: Studies on mechanisms of calcification. Johns Hopk. med. J. (Baltimore, U.S.A.) 127, 2–22 (1970)

    Google Scholar 

  • Bills, C. E., Eisenberg, H., Pallante, S. L.: Complexes of organic acids with calcium phosphate: The Von Kossa stain as a clue to the composition of bone mineral. Johns Hopk. med. J. (Baltimore, U.S.A.) 128, 194–207 (1971)

    Google Scholar 

  • Chappell, J. B., Perry, S. V.: The respiratory and adenosinetriphosphatase activities of skeletal muscle mitochondria. Biochem. J. 55, 586–595 (1953)

    Google Scholar 

  • Ebashi, S., Lipmann, F.: Adenosine triphosphate—linked concentration of calcium ions in a particulate fraction of rabbit muscle. J. Cell Biol. 14, 389–398 (1962)

    Google Scholar 

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

    Google Scholar 

  • Engel, A. G., Tice, L. W.: Cytochemistry of phosphatases of the sarcoplasmic reticulum. I. Biochemical studies. J. Cell Biol. 31, 473–487 (1966)

    Google Scholar 

  • Engel, W. K.: Adenosine triphosphatase of sarcoplasmic reticulum triads and sarcolemma identified histochemically. Nature (Lond.) 200, 588–589 (1963)

    Google Scholar 

  • Essner, E., Novikoff, A. B., Quintana, N.: Nucleoside phosphatase activities in rat cardiac muscle. J. Cell Biol. 25, 201–215 (1965)

    Google Scholar 

  • Gauthier, G. F.: On the localization of sarcotubular ATPase activity in mammalian skeletal muscle. Histochemie 11, 97–111 (1967)

    Google Scholar 

  • Gauthier, G. F., Padykula, H. A.: Cytochemical studies of adenosine triphosphatase activity in the sarcoplasmic reticulum. J. Cell Biol. 27, 252–260 (1965)

    Google Scholar 

  • Giacomelli, F., Bibbiani, C., Bergamini, E., Pellegrino, C.: Two ATPases in the sarcoplasmic reticulum of skeletal muscle fibres. Nature (Lond.) 213, 679–682 (1967)

    Google Scholar 

  • Gillespie, C. A., Simpson, D. R., Edgerton, V. R.: High glycogen content of red as opposed to white skeletal muscle fibres of guinea pigs. J. Histochem. Cytochem. 18, 552–558 (1970)

    Google Scholar 

  • Gordon, G. D., Price, H. M., Blumberg, J. M.: Electron microscopic localization of phosphatase activities within striated muscle fibers. Lab. Invest. 16, 422–435 (1967)

    Google Scholar 

  • Hasselbach, W.: ATP-driven active transport of Ca++ in the membranes of sarcoplasmic reticulum. Proc. roy. Soc. B 160, 501–504 (1964a)

    Google Scholar 

  • Hasselbach, W.: Relaxing factor and the relaxation of muscle. Progr. Biophys. molec. Biol. 14, 167–222 (1964b)

    Google Scholar 

  • Hasselbach, W., Makinose, M.: Die Calciumpumpe der “Erschlaffungsgrana” des Muskels und ihre Abhängigkeit von der ATP-Spaltung. Biochem. Z. 333, 518–528 (1961)

    Google Scholar 

  • Hasselbach, W., Makinose, M.: Über den Mechanismus des Calciumtransportes durch die Membranen des sarkoplasmatischen Reticulums. Biochem. Z. 339, 94–111 (1963)

    Google Scholar 

  • Hori, S. H., Takahashi, M.: An electron microscopic study of adenosine triphosphate—splitting enzyme in rat skeletal muscle by means of the section freeze substitution technique. Cytologia (Tokyo) 28, 331–341 (1963)

    Google Scholar 

  • Khan, M. A., Kakulas, B. A., Papadimitriou, J. M.: The effect of EDTA on the histochemical myofibrillar ATPase reaction. Presented at the 17th. Symposium of Gesellschaft für Histochemie in Bozen, Oct. 1974b. Acta histochem. (in press)

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

    Google Scholar 

  • Khan, M. A., Papadimitriou, J. M., Holt, P. G., Kakulas, B. A.: A calcium-citro-phosphate technique for the histochemical localization of myosin ATPase. Stain Technol. 47, 277–282 (1972b)

    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 (1973)

    Google Scholar 

  • 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 (1974a)

    Google Scholar 

  • Kielly, W. A.: Mitochondrial ATPase. In: Methods of enzymology, ed. S. P. Colowich and N. O. Kaplan, vol. 2, 593–595. New York: A.P. 1955

    Google Scholar 

  • MacLennan, D. H.: Purification and properties of an adenosine triphosphatase from sarcoplasmic reticulum. J. biol. Chem. 245, 4508–4518 (1970)

    Google Scholar 

  • Padykula, H. A., Gauthier, G. F.: Cytochemical studies of adenosine triphosphatases in skeletal muscle fibres. J. Cell Biol. 18, 87–107 (1963)

    Google Scholar 

  • Padykula, H. A., Herman, E.: The specificity of the histochemical method for adenosine triphosphatase. J. Histochem. Cytochem. 3, 170–183 (1955)

    Google Scholar 

  • Porter, K. R., Palade, G. E.: Studies on the endoplasmic reticulum. III. Its form and distribution in striated muscle cells. J. biophys. biochem. Cytol. 3, 269–300 (1957)

    Google Scholar 

  • Rostgaard, J., Behnke, O.: Fine structural localization of adenosine nucleoside phosphatase activity in the sarcoplasmic reticulum and the T system of rat myocardium. J. Ultrastruct. Res. 12, 579–591 (1965)

    Google Scholar 

  • Schulze, W., Wollenberger, A.: Elektronenmikroskopischer Nachweis von mitochondrialer Adenosin-triphosphatase Aktivität in tierischen Geweben. Histochemie 5, 417–429 (1965)

    Google Scholar 

  • Sommer, J. R., Spach, M. S.: Electron microscopic demonstration of adenosinetriphosphatase in myofibrils and sarcoplasmic membranes of cardiac muscle of normal and abnormal dogs. Amer. J. Path. 44, 491–505 (1964)

    Google Scholar 

  • Suko, J.: Alterations of Ca++ uptake and Ca++-activated ATPase of cardiac sarcoplasmic reticulum in hyper and hypothyroidism. Biochim. biophys. Acta (Amst.) 252, 324–327 (1971)

    Google Scholar 

  • Tice, L. W.: In situ studies of the sarcotubular system, p. 339–350. In: Factors influencing myocardial contractility, eds. R. D. Tanz, F. Kavaler and J. Roberts. New York: Academic Press 1967

    Google Scholar 

  • Tice, L. W., Engel, A. G.: Histochemical studies of a cation-sensitive adenosine triphosphatase of the sarcoplasmic reticulum. J. Cell Biol. 23, 97A (1964)

    Google Scholar 

  • Tice, L. W., Engel, A. G.: Cytochemistry of phosphatases of the sarcoplasmic reticulum. II. In situ localization of the Mg++ — dependent enzyme. J. Cell Biol. 31, 489–499 (1966)

    Google Scholar 

  • Wachstein, M., Meisel, E.: A histochemical demonstration of mitochondrial adenosine triphosphatases at a physiological pH with special reference to the demonstration of bile canaliculi. Amer. J. clin. path. 27, 13–23 (1957)

    Google Scholar 

  • Zebe, E.: Zur Lokalisation ATP-spaltender Reaktionen im „Sarcoplasmatischen Reticulum” quergestreifter Muskeln. Histochemie 5, 32–43 (1965)

    Google Scholar 

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Authors and Affiliations

  1. Department of Pathology, Perth Medical Centre, University of Western Australia, 6008, Shenton Park, 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. On the specificity of the histochemical technique for sarcoplasmic reticular adenosine triphosphatase: A light and electron microscopic study. Histochemistry 43, 101–111 (1975). https://doi.org/10.1007/BF00492439

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