, Volume 41, Issue 2, pp 119–131 | Cite as

Combined formaldehyde and acetyl chloride vapour condensation: A new fluorescence histochemical method for the demonstration of tryptophyl-peptide-containing endocrine cells of the hypophysis

  • Seppo Partanen
  • Leena Rechardt
  • Nils Bäck


The cells of the mammalian adenohypophysis store a substance, probably a tryptophyl-peptide, which exhibits formaldehyde-induced fluorescence (FIF). The fluorescence intensity and the number of cells in different parts of the adenohypophysis of human fetus or rat were studied after the following treatments: formaldehyde vapour alone, acidification of the formaldehyde-treated sections with hydrochloric acid or with glacial acetic acid, acetyl chloride vapour alone, as well as combined formaldehyde and acetyl chloride vapour.

The combined formaldehyde and acetyl chloride treatment induced an unexpectedly strong fluorescence in the pars intermedia of the rat, the fluorescence intensity, being 16 times that observed after formaldehyde treatment alone. In the pars distalis a moderate number of fluorescent cells was seen. After the acidification with hydrochloric acid the fluorescence intensity of the pars intermedia of the rat became 6 times stronger, but it did not cause the appearence of more fluorescent cells in the pars distalis. Acidification with glacial acetic acid had no effect on the fluorescence intensity. The intensity often began to increase only in the UV light, which is a new finding in the field of the, fluorescence histochemistry. The histochemical properties of the fluorogenic substance of the adenohypophysis of the human fetus and the rat were the same. The results of the combined formaldehyde and acetyl chloride treatment were easily reproducible, the fluorescence was strictly localized in the cytoplasm, and the microscopic structure of tissue was well preserved. The procedure is simple and easy to carry out. Thus, it offers good possibilities for the study of tryptophyl-peptides in various cells.


Fluorescence Intensity Hydrochloric Acid Glacial Acetic Acid Endocrine Cell Fluorescent Cell 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Axelsson, S., Björklund, A., Lindvall O.: Fluorescence histochemistry of biogenic monoamines: a study of the capacity of various carbonyl compounds to form fluorophores with biogenic monoamines in gas phase reactions. J. Histochem. Cytochem. 20, 435–444 (1972)Google Scholar
  2. Björklund, A., Falck, B.: Histochemical characterization of a tryptamine-like substance stored in cells of the mammalian adenohypophysis. Acta physiol. scand. 77, 475–489 (1969a)Google Scholar
  3. Björklund, A., Falck, B.: Pituitary monoamines, of the cat with special reference to the presence of an unidentified monoamine-like substance in the adenohypophysis. Z. Zellforsch. 93, 254–264 (1969b)Google Scholar
  4. Björklund, A., Falck, B., Lindvall, O., Svensson, L.-Å: New aspects on reaction mechanisms in the formaldehyde histofluorescence method for monoamines. J. Histochem. Cytochem. 21, 17–25 (1973)Google Scholar
  5. Björklund, A., Lindvall, O., Svensson, L.-Å: Mechanisms of fluorophore formation in the histochemical glyoxylic acid method for monoamines. Histochemie 32, 113–131 (1972)Google Scholar
  6. Björklund, A., Nobin, A., Stenevi, U.: Acid catalysis of the formaldehyde condensation reaction for a sensitive histochemical demonstration of tryptamines and 3-methoxylated phenylethylamines. J. Histochem. Cytochem. 19, 286–298 (1971)Google Scholar
  7. Corrodi H., Hillarp, N.-Å, Jonsson, G. Fluorescence methods for the histochemical demonstration of monoamines. 3. Sodium borohydride reduction of the fluorescent compounds as a specificity test. J. Histochem. Cytochem. 12, 582–586 (1964)Google Scholar
  8. Costa M, Eränkö, O.: Histochemical correlates, of cold-induced transsynaptic induction in the rat superior cervical ganglion. Histochem. J. 6, 35–53 (1974)Google Scholar
  9. Dahlström, A., Fuxe, K.: Monoamines and the pituitary gland. Acta endocr. (Kbh.) 51, 301–314 (1966)Google Scholar
  10. Eränkö, O.: Distribution of fluorescing islets, adrenaline and noradrenaline in the adrenal medulla of the hamster. Acta endocr. (Kbh.) 18, 174–179 (1955)Google Scholar
  11. Eränkö, O.: The practical histochemical demonstration of catecholamines by formaldehyde-induced fluorescence. J. roy. micr. Soc. 87, 259–276 (1967)Google Scholar
  12. Håkanson, R., Larsson, L.-I., Nobin, A., Sundler, F.: Tryptamine or tryptophyl peptides in endocrine cells of the mammalian adenohypophysis? J. Histochem. Cytochem. 20, 908–916 (1972)Google Scholar
  13. Håkanson, R., Melander, A., Owman, C., Sundler, F.: Depletion of secretory granules, calcitonin, and formaldehyde-ozone-induced fluorescence from cat thyroid C cells by vitamin D2 treatment. Histochemie 36, 89–96 (1973)Google Scholar
  14. Håkanson, R., Owman, C., Sundler, F.: Fluorescence histochemical and microspectrofluorometric evidence of tryptophyl peptides in thyroid C cells of cat and pig. J. Histochem. Cytochem. 20, 205–210 (1971)Google Scholar
  15. Håkanson, R., Sundler, F.: Formaldehyde condensation. A method for the fluorescence microscopic demonstration of peptides with NH2-terminal tryptophan residues. J. Histochem. Cytochem. 19, 477–482 (1971)Google Scholar
  16. Iturriza, F. C., Zieher, L. M., Ruager, J.: Detection of tryptamine in the pars distalis of the pituitary gland of the sheep. Acta endocr. Panam. 3, 121–127 (1972)Google Scholar
  17. Larsson, L.-I., Sundler, F., Håkanson, R., Rehfeld J. F., Stadil, F.: Immunofluorescent localization of gastrin in rabbit antropyloric mucosa to argyrophil cells exhibiting form-aldehyde-ozone-induced fluorescence. Histochemie 37, 81–87 (1973)Google Scholar
  18. Moriarty, G. C., Halmi, N. S.: Electron microscopic study of the adreno-corticotropin-producing cell with unlabeled antibody and soluble peroxidase-antiperoxidase complex J. Histochem. Cytochem. 20, 590–603 (1972)Google Scholar
  19. Partanen, S.: The acid-catalyzed formaldehyde-induced fluorescence in the hypophysis of the human fetus. Acta physiol. scand., in press (1974)Google Scholar
  20. Partanen, S., Hervonen, A.: Monoamine-containing structures in the hypothalamo-hypophyseal system in the human fetus. Z. Anat. Entwickl.-Gesch. 140, 53–60 (1973)Google Scholar
  21. Partanen, S., Rechardt, L.: Histochemically demonstrable monoamines in the pituitary gland and median eminence of the female rat during the postnatal development. Z. Zellforsch. 147, 41–57 (1973)Google Scholar
  22. Pearse, A. G. E.: Histochemistry. Theoretical and applied. London: J. and A. Churchill, Ltd. 1968Google Scholar
  23. Pearse, A. G. E., MacGregor, M. M.: Functional cytology of the pituitary gland. Ann. Rep. Brit. Emp. Cancer Camp. 2, 665–666 (1964)Google Scholar
  24. Pearse, A. G. E., Rost, F. W. D.: A new histochemical method for the, demonstration of tissue tryptophyl units by 2-acylation and ring closure., Histochem. J. 5, 577–578 (1973)Google Scholar
  25. Previro, A., Prota, G., Coletti-Previro, M. A.. C-Acylation of the tryptophan indole ring and its usefulness in protein chemistry. Biochim. biophys. Acta (Amst.) 285, 269–278 (1972)Google Scholar
  26. Tobyn, C., Leleux, P., Vanhaelst, L., Golstein, J., Herlant, M., Pasteels, J. L.: Immunohistochemical study of the human pituitary with anti-luteinizing hormone, anti-follicle stimulating hormone and anti-thyrotropin sera. Acta endocr. (Kbh.) 72, 625–642 (1973)Google Scholar
  27. Rost, F. W. D., Ewen, S. W. B.: New methods for the histochemical demonstration of catecholamines, tryptamines, histamine and other arylethylamines by acid-and aldehyde-induced fluorescence. Histochem. J. 3, 207–212 (1971)Google Scholar

Copyright information

© Springer-Verlag 1974

Authors and Affiliations

  • Seppo Partanen
    • 1
  • Leena Rechardt
    • 1
  • Nils Bäck
    • 1
  1. 1.Department of AnatomyUniversity of HelsinkiHelsinkiFinland

Personalised recommendations