, Volume 51, Issue 2–3, pp 141–152 | Cite as

Reduction and azo coupling of quinones

A histochemical study of human cutaneous melanin and adrenochrome
  • Ralph D. Lillie
  • Patricia T. Donaldson
  • Linda L. Vacca
  • Philip P. Pizzolato
  • Shitalkumar K. Jirge


Cutaneous melanin in formol fixed skin and adrenochrome in dichromate fixed monkey adrenal after adequate bisulfite or dithonite reduction were found to give definite azo coupling reactions. Weaker reactions were obtained on unreduced material, and these disappeared on ferric chloride oxidation. Both cutaneous melanin and adrenochrome appear to exist in a quinhydrone status. Prolongation of dichromate treatment weakens or abolishes azo coupling capacity of adrenochrome. The findings support the concept of quinonization and reduction to prevent and restore azo coupling of enterochromaffin cells and noradrenaline islets of the adrenal.

The most effective diazos for melanin werep-nitrodiazobenzene, fast black K and the diazosulfanilic acid, pH 1 pyronin B procedure, for adrenochrome. Diazosafranin and 2-chloro-4-nitrodiazobenzene were also useful. Blue and violet coupling products from toluidine blue and methylene violet RR fail to yield sufficient contrast to be convincing.


Bisulfite Toluidine Blue Ferric Chloride Dichromate Methylene Violet 
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  1. Barka, T., Anderson, P.J.: Histochemistry, theory, practice, and bibliography. New York: Hoeber Medical Div., Harper and Row Publishing Inc. 1963Google Scholar
  2. Barter, R., Pearse, A.G.E.: Detection of 5-hydroxytryptamine in mammalian enterochromaffin cells. Nature (Lond.)172, 810 (1953)Google Scholar
  3. Barter, R., Pearse, A.G.E.: Mammalian enterochromaffin as the source of serotonin (5-hydroxytryptamine). J. Path. Bact.69, 25–31 (1955)Google Scholar
  4. Benditt, E.P., Wong, R.E., Arase, M., Roeper, E.: 5-Hydroxytryptamine in mast cells. Proc. Soc. exp. Biol. (N.Y.)90, 303–304 (1955)Google Scholar
  5. Cohen, G.N.: Sur le premier produit de transformation de l'adrénochrome au cours de la mélanisation de l'adrénaline. C.R. Acad. Sci. (Paris)220, 796–797, 977–978 (1945)Google Scholar
  6. Cohen, G., Nisman, B.: Étude sur la mélanisation. IV. Structure des precurseurs des mélaninines. Régle et exception. Bull. Soc. Chim. biol. (Paris)29, 265–269 (1947)Google Scholar
  7. Coupland, R.E., Riley, J.F.: Mast cells and 5-hydroxytryptamine in precancerous mouse skin. Nature (Lond.)187, 1128–1129 (1960)Google Scholar
  8. Ganter, P., Jolles, G.: Histochimie, normale et pathologique. Paris: Gauthier-Villars 1969Google Scholar
  9. Gomori, G.: Microscopic histochemistry. Chicago: University Chicago Press 1952Google Scholar
  10. Heidenhain, R.: Untersuchungen über den Bau der Labdrüsen. Arch. mikr. Anat.6, 368–406 (1870)Google Scholar
  11. Hickinbottom, W.J.: Reactions of organic compounds, 3rd ed. London: Longmans Green 1957Google Scholar
  12. Lillie, R.D.: Histopathologic technic and practical histochemistry, 2nd ed. New York: Blakiston 1954Google Scholar
  13. Lillie, R.D.: The basophilia of melanins. J. Histochem. Cytochem.3, 453–454 (1955)Google Scholar
  14. Lillie, R.D.: Metal reduction reactions of the melanins: histochemical studies. J. Histochem. Cytochem.5, 325–333 (1957a)Google Scholar
  15. Lillie, R.D.: Trichoxanthin, the yellow granular pigment of guinea pig hair follicles and hair. J. Histochem. Cytochem.5, 346–353 (1957b)Google Scholar
  16. Lillie, R.D.: Metal chelate reaction of enterochromaffin. J. Histochem. Cytochem.9, 44–48 (1961a)Google Scholar
  17. Lillie, R.D.: Investigations on the structure of the enterochromaffin substance. J. Histochem. Cytochem.9, 184–191 (1961b)Google Scholar
  18. Lillie, R.D.: Histochemistry of melanins. Ch. 11. In: M. Wolman, Pigments in pathology. New York: Academic Press 1969Google Scholar
  19. Lillie, R.D.: Conn's biological stains, 9th ed. Baltimore: Williams and Wilkins 1977 (in press)Google Scholar
  20. Lillie, R.D., Burtner, H.J., Henson, J.P.G.: Diazo-safranin for staining enterochromaffin. J. Histochem Cytochem.1, 154–159 (1953)Google Scholar
  21. Lillie, R.D., Donaldson, P.T.: Histochemical azo coupling of protein histidine. Brunswik's nitration method. J. Histochem. Cytochem.20, 929–937 (1972)Google Scholar
  22. Lillie, R.D., Fullmer, H.M.: Histopathologic technic and practical histochemistry, 4th ed. New York-London: McGraw Hill 1976Google Scholar
  23. Lillie, R.D., Glenner, G.G.: Histochemical reactions of carcinoid tumors of the human gastrointestinal tract. Amer. J. Path.56, 623–651 (1960)Google Scholar
  24. Lillie, R.D., Palmer, R.W., Gutiérrez, A.: A diazosulfanilic acid azure A sequence for tissue protein. J. Histochem. Cytochem.21, 421–424 (1966)Google Scholar
  25. Lillie, R.D., Pizzolato, P.: Histochemical azo coupling reactions of the pigments of obstructive icterus and of hematoidin. I. Diazonium salts used. J. Histochem. Cytochem.17, 738–748 (1969)Google Scholar
  26. Lillie, R.D., Pizzolato, P., Vacca, L.L., Catalano, R.A., Donaldson, P.T.: Use of the diazosufanilic acid, pH 1 azure A sequence on adrenal medulla and the effects of prior oxidation and reduction on the reaction in several species. J. Histochem. Cytochem.21, 448–454 (1973a)Google Scholar
  27. Lillie, R.D., Pizzolato, P., Vacca, L.L., Catalano, R.A., Donaldson, P.T.: Histochemical azo coupling reactions: A catecholamine in enterochromaffin cells in place of, or in addition to 5-hydroxytryptamine. J. Histochem. Cytochem.21, 455–463 (1973b)Google Scholar
  28. Lison, L.: Sur quelques charactéristiques histochimiques d'un propigment mélanique. C.R. Soc. Biol. (Paris)106, 41–43 (1931a)Google Scholar
  29. Lison, L.: Recherches histochimiques sur les phénols et leurs derivés. Arch. Biol. (Paris)41, 343–436 (1931b)Google Scholar
  30. Lison, L.: Histochimie animale. Paris: Gauthier-Vielars 1936Google Scholar
  31. Lison, L.: Histochimie et cytochimie animales, 2 ed. Paris: Gauthier-Villars 1953Google Scholar
  32. Lison, L.: Histochimie et cytochimie animales, 3 ed. Paris: Gauthier-Villars 1960Google Scholar
  33. Lorincz, A.L.: Pigmentation. In: S. Rothman, Physiology and biochemistry of the skin. Ch. 22, pp. 515–563. Chicago: University Chicago Press 1954Google Scholar
  34. Meier, H.: Serotonin (5-hydroxytryptamine) content of canine mastocytoma. Proc. Soc. exp. Biol. (N.Y.)100, 815–817 (1959)Google Scholar
  35. Pearse, A.G.E.: Histochemistry, theoretical and applied. Boston: Little, Brown and Co. 1953Google Scholar
  36. Pearse, A.G.E.: Histochemistry, theoretical and applied, 3rd ed., Vol. 2. Edinburgh, London: Churchill, Livingstone 1972Google Scholar
  37. Raper, H.S.: The tyrosinase-tyrosine reaction. VI. Production from tyrosine of 5∶6-dihydroxyindole and 5∶6-dihydroxyindole-2-carboxylic acid, the precursors of melanin. Biochem. J.21, 89–96 (1927)Google Scholar
  38. Rice, W.G., Mitchener, J.: Histochemical evidence of 5-hydroxytryptamine in a dog mast cell tumor. Nature (Lond.)189, 767–768 (1961)Google Scholar
  39. Richter, D., Blaschko, H.: An oxidation product of adrenaline. J. chem. Soc.1937, 601–602Google Scholar
  40. Selye, H.: The mast cells. London: Butterworth 1965Google Scholar
  41. Thompson, S.W.: Selected histochemical and histopathological methods. Springfield, Ill.: C.C. Thomas 1966Google Scholar
  42. Vialli, M.: Considerazioni critiche sulla diagnosticabilita della presenza di catecolamine nelle cellule enterocromaffini. Acta histochem. (Jena)52, 88–99 (1975)Google Scholar

Copyright information

© Springer-Verlag 1977

Authors and Affiliations

  • Ralph D. Lillie
    • 1
  • Patricia T. Donaldson
    • 1
  • Linda L. Vacca
    • 1
  • Philip P. Pizzolato
    • 1
  • Shitalkumar K. Jirge
    • 1
  1. 1.Department of PathologyLouisiana State University Medical CenterNew OrleansUSA

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