Synopsis
Different grades of the colloid stabilizer, polyvinyl alcohol, used for protecting unfixed cryostat sections during cytochemical reactions, may have different effects on enzymatic activity. The influence of three grades of polyvinyl alcohol on the activities of ‘soluble’, membrane-bound and membrane-enclosed enzymes has been investigated in unfixed sections; the activities were measured microdensitometrically.
The largest molecular weight polyvinyl alcohol (G18/140, mol. wt. about 90 000) did not retain glucose-6-phosphate dehydrogenase activity in sections of rat liver even when used at the maximum convenient concentration (12%); G04/140 and M05/140 (molecular weights of 15 000 and 25 000 respectively) retained this soluble enzyme if used at concentrations of 30 and 20% respectively.
At these concentrations, lactate dehydrogenase activity was apparently decreased when G04/140 and M05/140 were used; this diminished activity has been shown to be due to the need to establish optimal concentrations of reactants for each grade of polyvinyl alcohol and for each reaction. When optimal concentrations of reactants were used, the activities of this enzyme in the presence of each grade of polyvinyl alcohol were identical.
The presence of any type of polyvinyl alcohol did not influence the activities of mitochondrial succinate dehydrogenase or of the smooth endoplasmic reticulum enzyme, Δ5,3β-hydroxysteroid dehydrogenase. However, the presence of polyvinyl alcohol improved the state of the section.
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References
Altman, F. P. (1971). The use of a new grade of polyvinyl alcohol for stabilising tissue sections during histochemical incubations.Histochemie 28, 236–42.
Altman, F. P. (1972). Quantitative dehydrogenase histochemistry with special reference to the pentose shunt dehydrogenases.Prog. Histochem. Cytochem.,4, No. 4.
Altman, F. P. &Chayen, J. (1965). Retention of nitrogenous material in unfixed tissue sections during incubation for histochemical demonstration of enzymes.Nature 207, 1205–6.
Altman, F. P. &Chayen, J. (1966). The significance of a functioning hydrogen transport system for the retention of ‘soluble’ dehydrogenases in unfixed sections.Jt R. Microsc. Soc. 95, 175–80.
Balogh, K., Jr. (1966). Histochemical demonstration of 3α-hydroxysteroid dehydrogenase activity.J. Histochem Cytochem. 14, 77–83.
Beyer, K. F. &Samuels, L. T. (1956). Distribution of steroid-3β-ol-dehydrogenase in cellular structures of the adrenal gland.J. biol. Chem. 219, 69–76.
Butcher, R. G. (1969). The influence of pharmacologically active substances in oxidative interactions between cytoplasm and mitochondria. Ph.D. Thesis, University of London.
Butcher, R. G. (1970). Studies on suceinate oxidation. 1. The use of intact tissue sections.Expl. Cell Res. 60, 54–60.
Butcher, R. G. &Chayen, J. (1968). The value of intact tissue sections for studying metabolic interactions between the cytoplasm and mitochondria.Expl. Cell Res. 49, 656–65.
Chayen, J., Bitensky, L. &Butcher, R. G. (1973).Practical Histochemistry. London: Wiley.
Dahl, H. A. &From, S. H. (1971). Some effects of polyvinyl alcohol and polyvinyl pyrrolidone on the activity of lactate dehydrogenase and its isoenzymes.Histochemie 25, 182–90.
Dahl, H. A. &Mellgren, S. I. (1970). The effect of polyvinyl alcohol and polyvinyl pyrrolidone on diffusion artifacts in lactate dehydrogenase activity.Histochemie 24, 354–70.
Henderson, B. (1976). Quantitative cytochemical measurement of glyceraldehyde 3-phosphate dehydrogenase activity.Histochemistry,48, 191–204.
Henderson, B. (1977). Quantitative cytochemical studies of rheumatoid arthritis. Ph. D. thesis, Brunel University.
Inano, H., Machino, A. &Tamaoki, B. (1969). Localisation of the Δ5-3β-hydroxysteroid dehydrogenase and 21 hydroxylase activities in smooth surfaced microsomes of adrenals.Steroids,13, 357–63.
Jacobsen, N. O. (1969). The histochemical localization of lactic dehydrogenase soenzymes in the rat nephron by means of an improved polyvinyl alcohol method.Histochemie,20, 250–65.
Kunze, K. D. (1967). Beltrag zur Histochemie der Laktatdehydrogenase.Histochemie 11 350–9.
Quattropani, S. L. &Weisz, J. (1973). Conversion of progesterone to estrone and estradiolin vitro by the ovary of the infantile rat in relation to the development of its interstitial tissue.Endocrinology,93, 1269–76.
Scott, J. E. (1974). The Feulgen reaction in polyvinyl alcohol or polyethylene glycol solution. ‘Fixation’ by excluded volume.J. Histochem. Cytochem. 22, 833–5.
Shannon, A. D. (1975). A post-coupling method for the demonstration ofN-acetyl-β D-glucosaminidase in unfixed frozen tissue sections.J. Histochem. Cytochem. 23, 424–30.
Stuart, J. &Skowron, P. N. (1968). A cytochemical study of marrow enzymes in negaloblastic anaemia.Br. J. Haemat. 15, 443–54.
Wenk, H. (1970). Beltrag zum histochemischen Nachweis lösclicher NAD-abhängiger Dehydrogenasen mittels PVA-Gelinkubation.Acta histochem. (Jena) 35, 380–91.
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Henderson, B., Loveridge, N. & Robertson, W.R. A quantitative study of the effects of different grades of polyvinyl alcohol on the activities of certain enzymes in unfixed tissue sections. Histochem J 10, 453–463 (1978). https://doi.org/10.1007/BF01003008
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DOI: https://doi.org/10.1007/BF01003008