Summary
An improved detection of activity of l-gulonolactone oxidase, which is responsible for the final oxidative step in the synthetic process of l-ascorbate from glucose in animals, was achieved using phenazine methosulfate and cyanide. Cold acetone fixation eliminated non-specific deposition of formazan on lipid droplets. The specificity of the method was tested and proven by a biological control, histochemical controls, inhibitors and activators. By application of the method, strong reactivity was found in the cytoplasm of centrilobular parenchymal cells of livers of the opossum, rat, ground squirrel and flying squirrel. Staining of dog liver was moderate and centrilobular. Prosimians were strongly positive: The centrilobular localization was found in the tree shrew and galago; slow lorises and some pottos showed strong reactivity in centrilobular cells and some peripheral cells as well. These prosimians seem to be able to synthesize l-ascorbate as many lower mammals are. On the contrary, true simians (i.e. the squirrel monkey, spider monkey, rhesus monkey and chimpanzee) were negative as guinea pigs were, suggesting their probable inability for l-ascorbate synthesis.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Brody, I. A., and W. K. Engel: Effects of phenazine methosulfate in histochemistry. J. Histochem. Cytochem. 12, 928–929 (1964).
Bublitz, C.: l-Gulono-γ-lactone oxidase and dehydrogenase. Biochim. biophys. Acta (Amst.) 48, 61–70 (1961).
—, and A. L. Lehninger: The role of aldonolactonase in the biosynthesis of l-ascorbic acid. Ann. N.Y. Acad. Sci. 92, 87–90 (1961).
Burns, J. J., P. Peyser, and A. Moltz: Missing step in guinea pigs required for the biosynthesis of l-ascorbic acid. Science 124, 1148–1149 (1956).
Burstone, M. S.: Hydrogen acceptors. In: Enzyme histochemistry and its application in the study of neoplasms, chap. 12/II, p. 477–487. New York and London: Academic Press 1962.
Caputto, R., R. E. Trucco, and A. E. Kitabchi: The interrelationship between gulonolactone oxidase and the process of lipid peroxidation. Ann. N.Y. Acad. Sci. 92, 79–86 (1961).
Chatterjee, I. B., G. C. Chatterjee, N. C. Ghosh, J. J. Ghosh, and B. C. Guha: Biological synthesis of l-ascorbic acid in animal tissues: Conversion of l-gulonolactone into l-ascorbic acid. Biochem. J. 74, 193–203 (1960).
— N. C. Kar, N. C. Ghosh, and B. C. Guha: Aspects of ascorbic acid biosynthesis in animals. Ann. N.Y. Acad. Sci. 92, 36–56 (1961).
Cohen, R. B.: Histochemical localization of l-gulonolactone oxidase activity in tissues of several species. Proc. Soc. exp. Biol. (N.Y.) 106, 309–311 (1961).
Day, P. L.: The nutritional requirements of primates other than man. Vitam. and Horm. 2, 71–105 (1944).
Dixon, M., and B. C. Webb: Enzyme inhibitors. In: Enzymes, 2nd ed., chap. VIII, p. 315–359. New York: Academic Press 1964.
Elliot, O., N. J. Yess, and D. M. Hegsted: Biosynthesis of ascorbic acid in the tree shrew and slow loris. Nature (Lond.) 212, 739–740 (1966).
Hart, K.: Über die experimentelle Erzeugung der Möller-Barlowschen Krankheit und ihre endgültige Identifizierung mit dem klassischen Skorbut. Virchows Arch. path. Anat. 208, 367–396 (1912).
Johnson, A. B.: The use of phenazine methosulfate for improving the histochemical localization of ketose reductase (l-iditol: NAD oxidoreductase, or sorbitol dehydrogenase). J. Histochem. Cytochem. 15, 207–215 (1967).
Kearney, E. B., and T. P. Singer: Studies on succinic dehydrogenase. I. Preparation and assay of the soluble dehydrogenase. J. biol. Chem. 219, 963–975 (1956).
Löw, H., B. Alm, and I. Vallin: The use of phenazine methosulfate in the study of oxidative phosphorylation. Biochem. biophys. Res. Commun. 14, 347–352 (1964).
Mathisen, J. S., and S. I. Mellgren: Some observation concerning the role of phenazine methosulfate in histochemical dehydrogenase methods. J. Histochem. Cytochem. 13, 408–409 (1965).
Miura, M., and N. Okabe: The antiscorbutic factor in commercially sterilized milk and Japanese green tea. Sci. Papers Inst. phys. chem. Res. (Tokyo) 20, 145–161 (1933).
Pearse, A. G. E., and R. Hess: Substantivity and other factors responsible for formazan patterns in dehydrogenase histochemistry. Experientia (Basel) 17, 136–141 (1961).
Rosa, C. G., and K.-C. Tsou: Use of tetrazolium compounds in oxidative enzyme histoand cytochemistry: Nature (Lond.) 192, 990–991 (1961).
Roy, R. N., and B. C. Guha: Species difference in regard to the biosynthesis of ascorbic acid. Nature (Lond.) 182, 319–320 (1958).
Shaw, J. H.: Vitamin C deficiency in the ringtail monkey. Fed. Proc. 8, 396 (1949).
Shimazono, N., and Y. Mano: Enzymatic studies on the metabolism of uronic and aldonic acids related to l-ascorbic acid in animal tissues. Ann. N.Y. Acad. Sci. 92, 91–104 (1961).
Talbot, F. B., W. J. Dodd, and H. O. Peterson: Experimental scorbutus and the Roentgen ray diagnosis of scorbutus. Boston med. surg. J. 169, 232–239 (1913).
Tomlinson, T. H., Jr.: Oral pathology in monkeys in various experimental dietary deficiencies. Publ. Hlth Rep. (Wash.) 54, 431–439 (1939).
Winkelman, J., and A. L. Lehninger: Aldono- and uronolactonases of animal tissues. J. biol. Chem. 233, 794–799 (1958).
Author information
Authors and Affiliations
Additional information
Visiting scientist from the Department of Anatomy, Tokyo Medical and Dental University, Tokyo, Japan.
T. R. Shanthaveerappa in previous publications, also fellow, Department of Anesthesiology, Emory University.
Rights and permissions
About this article
Cite this article
Nakajima, Y., Shantha, T.R. & Bourne, G.H. Histochemical detection of l-gulonolactone: phenazine methosulfate oxidoreductase activity in several mammals with special reference to synthesis of vitamin C in primates. Histochemie 18, 293–301 (1969). https://doi.org/10.1007/BF00279880
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00279880