The precipitation patterns of the following ultracytochemical methods in rat muscle cells were compared and examined critically: the potassium pyroantimonate method for calcium demonstration; the calcium phosphate technique for the Ca2+ — ATPase reaction; the formazan reaction for the demonstration of creatine kinase activity (all performed on heart muscle); and the lead phosphate technique for the Mg2+ — ATPase reaction in skeletal muscle. Using X-ray microanalysis, it was found that the antimonate precipitate contains only calcium as the precipitated ion in the vast majority of cases. Most probably it consists of pure calcium pyroantimonate. However, in myocytes showing the well-established precipitation pattern, the concentration of calcium was estimated to be about two orders of magnitude higher than the native concentration of total intracellular calcium. It is concluded that calcium ions diffuse freely from the extracellular space and from adjacent cells into cells containing antimonate and are precipitated mostly at sites where heterogeneous nucleation is facilitated by intracellular catalysts (biopolymers).
As shown by the similar precipitation patterns for the four reactions compared, these catalysts are not specific to any of these reactions and are most probably neither calcium-binding sites nor sites of any one of the enzymes examined in the native cell.
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AGUAS, A. P. & NICKERSON, P. A. (1981) Distribution of Ca differs in relaxed and contracted myocardial cells of the rat.Cell Tiss. Res. 221, 295–302.
BORGERS, H., THONE, F., VERHEYEN, A. & TER KEURS, H. E. D. J. (1984) Localization of calcium in skeletal and cardiac muscle.Histochem, J. 16, 295–301.
DE JONG, A. S. H. (1982) Mechanisms of metal-salt methods in enzyme cytochemistry with special reference to acid phosphatase.Histochem. J. 14, 1–33.
DICULESCU, I., IONESCU, N. & POPESCU, L. M. (1978) Calcium immobilization in mitochondria of failing myocardium: X-ray microanalytical evidence.J. mol. cell. Cardiol. 10, 1141–51.
DUX, E., TOTH, I., DUX, L. & JOO, F. (1978) The localization of Ca by X-ray microanalysis in myopathic muscle fibers.Histochemistry 56, 3–4.
FARBER, J. L., CHIEN, K. R. & MITTNACHT, S. (1981) The pathogenesis of irreversible cell injury in ischemia.Am. J. Path. 102, 271–81.
FARQUHAR, M. G. & PALADE, G. E. (1966) Adenosine triphosphatase localization in amphibian epidermis.J. Cell Biol. 30, 359–79.
FUJIMOTO, K. & OGAWA, K. (1982) Enzyme cytochemical study of rat cardiac muscle. II. Ca2+ — ATPase and ouabain-sensitive K+ — dependentp-nitrophenylphosphatase.Acta histochem. cytochem. 15, 338–54.
KOMNICK, H. (1962) Elektronenmikroskopische Lokalisation von Na+ und Cl− in Zellen und Geweben.Protoplasma 55, 414–8.
LEGATO, M. J. & LANGER, G. A. (1969) The subcellular localization of calcium ions in mammalian myocardium.J. Cell Biol. 41, 401–23.
MYKLEBUST, R., JUSTESEN, N. P. & SAETERSDAL, T. S. (1975) Troponin bound calcium in electron micrographs of myocardial cells as demonstrated by the potassiumantimonate technique.Cell Tiss. Res. 162, 323–39.
PADYKULA, H. A. & HERMAN, E. (1955) The specifity of the histochemical method for adenosine triphosphatase.J. Histochem. Cytochem. 3, 167–70.
PUNKT, K., SCHIPPEL, G., WELT, K. & SCHIPPEL, K. (1981) Ultrahistochemischer Nachweis der ATPase-Aktivität am Rattenherzen in der postnatalen Entwicklung. 10. Tagung Elektronenmikroscopie Leipzig, p. 117.
SAETERSDAL, T. S., MYKLEBUST, R., JUSTESEN, N. P. B. & OLSEN, W. (1974) Ultrastructural localization of calcium in the pigeon papillary muscle as demonstrated by cytochemical studies and X-ray microanalysis.Cell Tiss. Res. 155, 57–74.
SCHULZE, W. & WOLLENBERGER, A. (1971) Cytochemistry of membrane-bound ATPase system of cardiac muscle.Meth. Achievm. exp. Path. 5, 347–83.
SHAROV, V. G., SAKS, V. A., SMIRNOV, V. N. & CHZOV, E. I. (1977) An electron miscroscopic histochemical investigation of the localization of kreatine phosphokinase in heart cells.Biochem. biophys. Acta 468, 495–501.
SIMSON, J. A. V. & SPICER, S. S. (1975) Selective subcellular localization of cations with variants of the potassium pyroantimonate technique.J. Histochem. Cytochem. 23, 575–98.
SOMLYO, A. P. & SHUMAN, H. (1982) Electron probe and electron energy loss analysis in biology.Ultramicroscopy 8, 219–34.
UENO, S., MAYAHARO, H., TSUKAHARA, J. & OGAWA, K. (1980) Ultrahistochemical localization of ouabainsensitive, potassium-dependentp-Nitrophenylphosphatase activity in guinea pig retina.Acta histochem. cytochem. 13, 679–94.
VAN DUIJN, P. & VAN DER PLOEG, M. (1980) Microscopic cytochemistry as matrix chemistry. InTrends in Enzyme Histochemistry and Cytochemistry Ciba Foundation Symposium73, pp. 209–29. Amsterdam: Elsevier, North Holland.
VAN IREN, F. & VAN DER SPIEGEL, A. (1975) Subcellular localization of inorganic ions in plant cells byin vivo precipitations.Science 18, 1210–1.
VAN IREN, F, VAN ESSEN-JOOLEN, L., VAN DER DIUN-SCHOUTEN, P., VAN DER SLUIJS, P. B., & DE BRUIJN, W. C. (1979) Sodium and calcium localization in cells and tissues by precipitation with antimonate: A quantitative study.Histochemistry 63, 273–94.
YEH, B. K. (1973) Localization of calcium antimonate in the atrial and ventricular muscle fibers of the cat heart.J. mol. cell. Cardiol. 5, 351–8.
VON ZGLINICKI, T. (1983) Quantitative Röntgenmikroanalyse biologischer Ultradünnschnitte mit Aluminium-Kohle-Aufdampfschichten als Standards.Acta histochem. 72, 195–201.
VON ZGLINICKI, T., BIMMLER, M. & PURZ, H.-J. (1986) Fast cryofixation technique for X-ray microanalysis.J. Microsc.,141, 1.
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Von Zglinicki, T., Punkt, K. Localization artefacts in ultracytochemical ion precipitation reactions. Histochem J 18, 29–35 (1986). https://doi.org/10.1007/BF01676195
- Creatine Kinase
- Calcium Phosphate
- Precipitation Pattern
- Creatine Kinase Activity