Summary
The use of the Ca-phosphate-method for the determination of ATP-ase pH 7.5 is reported. The results obtained in experimental myocardial infarction, autolysis, and in other lesion-producing mechanisms suggest, that the activity in question seems to be a mitochondrial ATP-ase, which by uncoupling the oxidative phosphorylation gives rise to an increased activity, that is detectable by histochemical means. It is pointed out, that the fixation of the sections is of importance. Our findings suggest that under certain conditions the Ca-phosphate-method is suitable for the determination of ATP-ase in a low pH range.
Zusammenfassung
Es wird über die Anwendung der Ca-Phosphatmethode zur histochemischen Darstellung der ATPase pH 7,5 berichtet. Ergebnisse beim experimentellen Herzinfarkt, bei der Autolyse sowie anderen Schädigungsmechanismen deuten darauf hin, daß es sich um den Nachweis einer mitochondrialen ATPase handelt, die bei Entkopplung der oxydativen Phosphorylierung der Anlaß für eine histochemisch nachweisbare Aktivitätssteigerung ist. Es wird auf die Bedeutung einer vorherigen Fixierung der Schnitte hingewiesen. Unsere Befunde lassen die Annahme zu, daß unter gewissen Voraussetzungen auch die Ca-Phosphatmethode zum Nachweis der ATPase im niederen pH-Bereich geeignet ist.
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Literatur
Barden, H., S. S. Lazarus: Histochemical characteristics of adenosine triphosphate dephosphorylating enzymes in rabbit pancreas. J. Histochem. Cytochem. 11, 578–589 (1963).
Bradshaw, Marie, M. Wachstein, J. Spence, and J. M. Elias: Adenosine triphosphatase activity in melanocytes and epidermal cells of human skin. J. Histochem. Cytochem. 11, 465–473 (1963).
Chauncey, H. H., J. H. Kronman, and M. A. Levitt: Preservation of total, lyo- and desmoenzyme activity in mammalian salivary glands following chloral hydrate formalin fixation. J. Histochem. Cytochem. 12, 647–653 (1964).
Cooper, C., and A. L. Lehniniger: Oxidative phosphorylation by an enzyme complex from extracts of mitochondria. IV. Adenosintriphosphatase activity. J. biol. Chem. 224, 547–560 (1957).
Danielli, J. F.: A critical study of techniques for determining the cytological position of alkaline phosphatase. J. exp. Biol. 22, 110–117 (1946).
Deimling, O. H. v.: Die Darstellung phosphatfreisetzender Enzyme mittels SchwennetallSimultan-Methoden. Histochemie 4, 48–55 (1964).
Dianzani, M. U., and S. Scuro: The effects of some inhibitors of oxidative phosphorylation on the morphology and enzymic activities of mitochondria. Biochem. J. 62, 205–215 (1956).
Eder, M.: Histochemische Fermentnachweise. Ihre Bedeutung in der Pathologie. Verh. dtsch. Ges. Path. 42, 374–391 (1958).
Essner, E., A. B. Novrkoff, and B. Masek: Adenosintriphosphatase and 5-Nucleotidase activities in the plasma membrane of liver cells as revealed by electron microscopy. J. biophys. biochem. Cytol. 4, 711–716 (1958).
Gallagher, C. H., J. D. Judah, and K. R. Rees: Enzyme changes during liver autolysis. J. Path. Bact. 72, 247–256 (1956).
Gauthier, G. F., and Helen A. Padykula: Cytochemical studies of the ATPases of skeletal muscle fibres. J. Histochem. Cytochem. 10, 661–662 (1962).
Gomori, G.: Pitfalls in histochemistry. Ann. N.Y. Acad. Sci. 50, 968–978 (1950).
—, E. P. Benditt: Precipitation of calcium phosphate in the histochemical method for phosphatase. J. Histochem. Cytochem. 1, 114–122 (1953).
Greville, D. G. and D. M. Needham: Effect of 2∶4-dinitrophenol and phenylmercuric acetate on enzymic activity of myosin. Biochim. biophys. Acta (Amst.) 16, 284–285 (1955).
Hannibal, M. J. and M. M. Nachlas: Further studies on the lyo and desmo components of several hydrolytic enzymes and their histochemical significance. J. biophys. biochem. Cytol. 5, 279–288 (1959).
Harman, J. W., and A. Kitiyakara: Studies on mitochondria. VI, The relationship between the structure, osmotic reactivity and ATPase activity of mitochondria from pigeon skeletal muscle. Exp. Cell Res. 8, 411–435 (1955).
Hecht, A.: Fermenthistochemische Frühveränderungen beim experimentellen Herzinfarkt. Virchows Arch. path. Anat. 337, 414–424 (1964).
Holzner, J. H., u. F. Kaufman: Der Einfluß der postmortalen Autolyse auf die histochemisch darstellbare Adenosintriphosphatase-(ATPase-)Aktivität. Path. et Microbiol. (Basel) 25, 193–203 (1962).
Hori, S. H. and J. P. Chang: Histochemical study of adenosine triphosphatase in cytoplasm. J. Histochem. Cytochem. 11, 71–79 (1963).
Kaltenbach, J. C., and J. W. Harman: VII. The relationship between the control of structure and the dinitrophenol stimulation of adenosinetriphosphatase in liver mitochondria. Exp. Cell Res. 8, 435–452 (1955).
Kielley, W. W., and O. Meyerhof: Studies on adenosinetriphosphatase of muscle. II. A new magnesium-activated adenosinetriphosphatase. J. biol. Chem. 176, 591–601 (1948).
Klemperer, H. G.: ATPase activity of rat liver mitochondria. Biochim. biophys. Acta (Amst.) 23, 404–412 (1957).
Lardy, H. A., and H. Wellman: The catalytic effect of 2,4-dinitrophenol on adenosinetriphosphate hydrolysis by cell particles and soluble enzymes. J. biol. Chem. 201, 357–370 (1953).
Lazarus, S. S., and H. Barden: Histochemistry and electron microscopy of mitochondrial adenosinetriphosphatase. J. Histochem. Cytochem. 10, 285–293 (1962).
Lehntnger, A. L.: Water uptake and extrusion by mitochondria in relation to oxidative phosphorylation. Physiol. Rev. 42, 467–517 (1962).
Merker, H.-J., J. W. Wedell u. D. Neubert: Biochemische und strukturelle Veränderungen an den Zellorganellen der Leber nach vollständiger Kreislaufunterbrechung. NaunynSchmiedebergs Arch. exp. Path. Pharmak. 249, 85–116 (1964).
Muscatello, U., E. Andersson-Cedergren, G. F. Azzone, and A. von der Decken: The sarcotubular system of frog skeletal muscle. J. biophys. biochem. Cytol. 10, Suppl. 201–218 (1961).
Myers, D. K., and E. C. Slater: The enzymic hydrolysis of adenosine triphosphate by liver mitochondria. 1. Activities at different pH values. Biochem. J. 67, 558–572 (1957).
— The enzymic hydrolysis of adenosine triphosphate by liver mitochondria. 2. Effect of inhibitors and added cofactors. Biochem. J. 67, 572–579 (1957).
Nachlas, M. M., W. Prinn, and A. M. Seligman: Quantitative estimation of lyo- and desmoenzymes in tissue sections with and without fixation. J. biophys. biochem. Cytol. 2, 487–502 (1956).
Niles, N. R., J. Chayen, G. J. Cunningham and L. Bitensky: The histochemical demonstration of adenosine triphosphatase activity in myocardium. J. Histochem. Cytochem. 12, 740–743 (1964).
Novikoff, A. B., D. H. Hausman and E. Podber: The localization of adenosinetriphosphatase in liver: in situ staining and cell fraction studies. J. Histochem. Cytochem. 6, 61–71 (1958).
Padykula, Helen A. and G. F. Gauthier: Cytochemical studies of adenosine triphosphatase in skeletal muscle fibres. J. Cell. Biol. 18, 87–107 (1963).
—, Edith Herman: Factors affecting the activity of adenosinetriphosphatase and other phosphatases as measured by histochemical techniques. J. Histochem. Cytochem. 3, 161–169 (1955).
— The specifity of the histochemical method for adenosine triphosphatase. J. Histochem. Cytochem. 3, 170–195 (1955).
Pearse, A. G. E. and J. L. Reis: The histochemical demonstration of a specific phosphatase (5-Nucleotidase). Biochem. J. 50, 534–536 (1952).
Perry, S. V.: The adenosinetriphosphatase activity of lipoprotein granules isolated from skeletal muscle. Biochim. biophys. Acta (Amst.) 8, 499–509 (1952).
Potter, V. R., P. Siekevitz and H. Simonson: Latent adenosinetriphosphatase activity in resting rat liver mitochondria. J. biol. Chem. 205, 893–908 (1953).
Purvis, J. L., and E. C. Slater: The effect of magnesium on oxidative phosphorylation and mitochondrial adenosine triphosphatase. Exp. Cell Res. 16, 109–117 (1959).
Racker, E.: Mechanisms of synthesis of adenosine triphosphate. Advanc. Enzymol. 23, 323–399 (1961).
Scarpelli, D. G., and A. G. E. Pearse: Physical and chemical protection of cell constituents and the precise localization of enzymes. J. Histochem. Cytochem. 6, 369–376 (1958).
Schreiber, G., u. H. Simon: Kritische Untersuchungen zum histochemischen Nachweis der Phosphatasen sowie deren Beeinflußbarkeit in vitro und in vivo. Histochemie 4, 252–260 (1964).
Stein, J. M., and H. A. Padykula: Histochemical classification of individual skeletal muscle fibres of the rat. Amer. J. Anat. 110, 103–123 (1962).
Vorbrodt, A.: The effect of some inhibitors of oxidative phosphorylation on the histochemically demonstrable phosphatases. Exp. Cell Res. 12, 154–162 (1957).
Wachstein, M., M. Bradshaw, and J. M. Ortiz: Histochemical demonstration of mitochondrial adenosine triphosphatase activity in tissue sections. J. Histochem. Cytochem. 10, 65–74 (1962).
—, E. Meisel: Histochemistry of hepatic phosphatases at physiologic pH with special reference to the demonstration of bile canaliculi. Amer. J. clin. Path. 27, 13–23 (1957).
—, E. Meisel and A. Niedzwiedz: Histochemical demonstration of mitochondrial adenosine triphosphatase with the lead-adenosine triphosphate technique. J. Histochem. Cytochem. 8, 387–388 (1960).
Wegmann, R., et Z. Bankowski: Différenciation histochimique de quatre groupes d'Adenosine triphosphatases selon Slater. Ann. Histochim. 5, 121–141 (1960).
Witter, R. F., M. L. Watson, and M. A. Cottone: Morphology and ATPase of isolated mitochondria. J. biophys. biochem. Cytol. 1, 127–138 (1955).
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Auszugsweise vorgetragen auf dem II. Internationalen Kongreß für Histo- und Cytochemie, Frankfurt a. M. vom 16.-21. 8. 64.
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Hecht, A. Erfahrungen in der Anwendung der Ca-Phosphatmethode bei PH 7,5 zum Histochemischen Nachweis der Adenosintriphosphatase. Histochemie 5, 116–124 (1965). https://doi.org/10.1007/BF00285503
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DOI: https://doi.org/10.1007/BF00285503