Summary
Final reaction product formation was recorded microphotometrically for succinate dehydrogenase in cross-sectioned muscle fibres at initial rate conditions and during prolonged incubations. Incubations with gel films and aqueous reaction medium both showed a decline of reaction rates. Maximum reaction rates could only be determined at initial rate conditions during the first minute of the incubation. Reaction rates recorded in different areas of the same tissue section were found to change with time to different degrees. From these results it was concluded that quantitative histochemical measurements of enzyme reactionsin situ can only be valid if measured under initial maximum velocity conditions.
Similar content being viewed by others
References
Bitensky, L. (1980) Microdensitometry. InTrends in Enzyme Histochemistry and Cytochemistry. Ciba Foundation Symposium, Vol. 73 (new series), pp. 181–202. Amsterdam, Oxford, New York: Excerpta Medica.
Butcher, R. G. (1978) The measurement in tissue sections of the two formazans derived from Nitroblue Tetrazolium in dehydrogenase reactions.Histochem. J. 10, 739–44.
Nolte, J. &Pette, D. (1971) Quantitative microscope-photometric determination of enzyme activities in cryostat sections. InRecent Advances in Quantitative Histo- and Cytochemistry (edited byDubach, U. C. &Schmidt, U.), pp. 54–60. Bern, Stuttgart, Vienna: Hans Huber.
Nolte, J. &Pette, D. (1972) Microphotometric determination of enzyme activity in single cells in cryostat sections. I. Application of the gel film technique to microphotometry and studies on the intralobular distribution of succinate dehydrogenase and lactate dehydrogenase activities in rat liver.J. Histochem. Cytochem. 20, 567–76.
Pette, D. &Brandau, H. (1962) Intracellular localization of glycolytic enzymes in cross-striated muscle ofLocusta migratoria.Biochem. biophys. Res. Commun. 9, 367–70.
Pette, D., Wasmund, H., &Wimmer, M. (1979) Principle and method of kinetic microphotometric enzyme activity determinationin situ.Histochemistry 64, 1–10.
Pette, D. &Wimmer, M. (1979) Kinetic microphotometric activity determination in enzyme containing gels and model studies with tissue sections.Histochemistry 64, 11–22.
Pette, D. &Wimmer, M. (1980) Microphotometric determination of enzyme activities in cryostat sections by the gel film technique. InTrends in Enzyme Histochemistry and Cytochemistry. Ciba Foundation Symposium Vol. 73 (new series), pp. 121–134. Amsterdam, Oxford, New York:Excerpta Medica.
Pette, D., Wimmer, M. &Nemeth, P. (1980) Do enzyme activities vary along muscle fibres?Histochemistry 67, 225–31.
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, 9–19.
Spamer, C. &Pette, D. (1980) Metabolic subpopulations of rabbit skeletal muscle fibres. InPlasticity of Muscle (edited byPette, D.), pp. 19–30. Berlin, New York: de Gruyter.
Stoward, P. J. (1980) Criteria for the validation of quantitative histochemical enzyme techniques. InTrends in Enzyme Histochemistry and Cytochemistry. Ciba Foundation Symposium, Vol. 73 (new series), pp. 11–31. Amsterdam, Oxford, New York: Excerpta Medica.
Wimmer, M. &Pette, D. (1979) Microphotometric studies on intraacinar enzyme distribution in rat liver.Histochemistry,64, 23–33.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Pette, D. Microphotometric measurement of initial maximum reaction rates in quantitative enzyme histochemistryin situ . Histochem J 13, 319–327 (1981). https://doi.org/10.1007/BF01006885
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01006885