Synopsis
A fluorescence metachromatic modification of the ‘masked basophilia’ method is described. It is based on the acridine dye Coriphosphine O. Excitation and emission spectra of green (orthochromatic) and red (metachromatic) fluorescent tissue components are presented.
When the method is applied to suitably fixed sections, metachromasia is demonstrable in cells of the polypeptide hormone-secreting APUD series, and in a few other situations.
The view that side-chain carboxyl groups are demonstrated by the masked basophilia technique is considered to be accurate but inadequate. It is proposed that the technique, and its fluorescence modification, are influenced more by secondary than by primary protein structure. In particular, it is suggested that the conformation of the protein precursors of polypeptide hormones, in the storage granules of endocrine cells, is predominantly random-coil.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Barrnett, R. j. &Seligman, A. m. (1958). Histochemical demonstration of proteinbound alpha-acylamido carboxyl groups.J. biophys. biochem. Cytol. 4, 169–76.
Blaschko, H., Hagen, J. m. &Hagen, P. (1957). Mitochondrial enzymes and chromaffin granules.J. Physiol., Lond. 139, 316–22.
Carvalheira, A. f., Welsch, U. &Pearse, A. g. e. (1968). Cytochemical and ultrastructural observations on the argentaffin and argyrophil cells of the gastrointestinal tract in mammals, and their place in the APUD series of polypeptide-secreting cells.Histochemie 14, 33–46.
Cavallero, C., Solcia, E. &Sampietro, R. (1967). Cytology of islet tumours and hyperplasias associated with the Zollinger-Ellison syndrome.Gut 8, 172–7.
Chibnall, A. c., Haselbach, C., Mangan, J. l. &Rees, M. w. (1958) Studies on the amide and C-terminal residues in proteins. 5. Estimation of asparagine and glutamine residues.Biochem. J. 68, 122–8.
Dean, C. r. &Hope, D. b. (1968). The isolation of neurophysin-I &-II from bovine pituitary neurosecretory granules separated on a large scale from other subcellular organelles.Biochem. J. 106, 565–73.
Francois, C. j. &Glimcher, M. j. (1967). The isolation and amino-acid composition of the α-chains of chicken bone collagen.Biochim. biophys. Acta 133, 91–6.
Gouinlock, E. v., Flory, P. j. &Scheraga, H. a. (1955). Molecular configuration of gelatin.J. Polym. Sci. 16, 383–95.
Haitinger, M. (1938). Fluoreszenz-Mikroskopie. Leipsig: Akademie-Verlag (reprinted with additions, ed. Eisenbrand and Werth, 1959).
Hill, R. l. (1965). Hydrolysis of Proteins.Adv. Protein Chem. 20, 37–107.
Hillson, P. j. &Mckay, R. b. (1966). Metachromasy.Nature, Lond. 210, 297.
Hopwood, D. (1967). Adrenal medullary basophilia in ox, pig, and sheep. A histochemical, immunohistochemical and cell fractionation study.Histochemie II, 268–79.
Karnovsky, M. j. &Fasman, G. d. (1960). A histochemical method for distinguishing between side-chain and terminal carboxyl groups of proteins.J. Histochem. Cytochem. 8, 326–7.
Kopple, K. d. (1966).Peptides and Amino Acids. New York: Benjamin, p. 82.
Kuyper, C. m. a. (1962). The metachromasia of fluorescence of acridine dyes.Histochemie 3, 46–53.
Manocchio, I. (1960). Metachromatische Färburg der A-Zellen in Pancreasinseln von Canis Familiaris.Zbl. allg. Path. path. Anat. 101, 1–4.
Manocchio, I. (1964). Metachromasia e basofilia delle cellule insulari alfa nel pancreas di mammiferi dopo metilzione e demetilazione.Arch. vet. ital. 15 3.
Mcguigan, J. e. (1968). Gastric mucosal intracellular localization of gastrin by immunofluorescence.Gastroenterology 55, 315–27.
Pearse, A. g. e. (1966a). Common cytochemical properties of cells producing polypeptide hormones, with particular reference to calcitonin and the thyroid C cells.Vet. Rec. 79, 587–90.
Pearse, A. g. e. (1966b). The cytochemistry of the thyroid C cells and their relationship to calcitonin.Proc. R. Soc. B. 164, 478–87.
Pearse, A. g. e. (1968a). Common cytochemical and ultrastructural characteristics of cells producing polypeptide hormones (the APUD series) and their relevance to thyroid and ultimobranchial C cells and calcitonin.Proc. R. Soc. B. 170, 71–80.
Pearse, A. g. e. (1968b). The thyroid parenchymatous cells of Baber, and the nature and function of their C cell successors in thyroid, parathyroid and ultimobranchial bodies. InCalcitonin (Proc. Symp. Thyrocalcitonin and the C cells). London: Heinemann, pp. 98–109.
Pearse, A. g. e. (1969). Cytochemical evidence for random coil conformation of polypeptide hormone precursor protein in endocrine cells.Nature, Lond. 221, 1210–11.
Pearse, A. g. e. &Rost, F. w. d. (1969). A microspectrofluorimeter with epiillumination and photon counting.J. Microscopy 89, 294–311.
Rigler, R. (1966). Microfluorometric characterization in intracellular nucleic acids and nucleoproteins by Acridine Orange.Acta physiol. scand. 67, Suppl. 267.
Schiebe, G. &Zanker, V. (1962). Uber die Metachromasie der Absorption und Fluoreszenz von Acridinefarbstoffen.Histochemie,3, 122–6.
Smith, A. d. &Winkler, H. (1967). Purification and properties of an acidic protein from chromaffin granules of bovine adrenal medulla.Biochem. J. 103, 483–92.
Solcia, E. &Sampietro, R. (1965a). Osservazioni sulla Basofilia delle Cellule Argentofile e di quelle Enterochromaffini.Riv. Istochim. norm. patol. 9, 265–78.
Solcia, E. &Sampietro, R. (1965b). Cytologic observations on the pancreatic islets with reference to some endocrine-like cells of the gastro-intestinal mucosa.Z. Zellforsch. mikrostr. Anat. 68, 689–98.
Solcia, E. &Sampietro, R. (1968). New methods for staining secretory granules and 5-hydroxytryptamine in the thyroid C cells. InCalcitonin (Proc. Symp. Thyrocalcitonin and the C cells). London: Heinemann, pp. 127–32.
Solcia, E., Vassallo, G. &Capella, C. (1968). Selective staining of endocrine cells by basic dyes after acid hydrolysis.Stain Technol. 43, 257–63.
Solcia, E., Vassallo, G. &Sampietro, R. (1967). Endocrine cells in the antropyloric mucosa of the stomach.Z. Zellforsch. mikrostr. Anat. 81, 474–86.
Stone, A. l. (1967). Aggregation of cationic dyes on acid polysaccharides. II. Quantitative parameters of metachromasy.Biochim. biophys. Acta 148, 193–206.
Stone, A. l. &Bradley, D. f. (1967). Aggregation of cationic dyes on acid polysaccharides. I. Spectrophotometric titration with Acridine Orange and other metachromatic dyes.Biochim. biophys. Acta 148, 172–92.
Stoward, P. j. (1967). Studies in fluorescence histochemistry. I. The demonstration of sulphomucins.Jl R. microsc. Soc. 87, 215–35.
Stoward, P. j. &Burns, J. (1967). Studies in fluorescence histochemistry. IV. The demonstration of the C-terminal carboxyl group in proteins.Histochemie 10, 230–3.
Stoward, P. j. &Burns, J. (1968). Some evidence confirming the specificity of Barrnett and Seligman's technique for demonstrating side-chain carboxyl groups in proteins.Histochemie 13, 7–10.
West, S. s. (1965) Fluorescence microspectroscopy of mouse leukocytes supravitally stained with acridine orange.Acta histochem. Suppl. 6, 135–56.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Bussolati, G., Rost, F.W.D. & Pearse, A.G.E. Fluorescence metachromasia in polypeptide hormone-producing cells of the APUD series, and its significance in relation to the structure of the precursor protein. Histochem J 1, 517–530 (1969). https://doi.org/10.1007/BF01012858
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF01012858