Summary
Serotonin immunoreactive material was localized to rat enterochromaffin cells (EC cells) at the subcellular level using antibodies to serotonin (5-HT) raised in rabbits. Ultrathin sections from paraformaldehyde fixed plastic embedded tissues were directly labelled with the 5-HT antiserum, using the protein A-gold technique to visualize the immunoreaction. The 5-HT immunoreactivity (5-HT-IR) in the rat gastrointestinal mucosa was exclusively localized to epithelial EC cells with a low background over other epithelial non-enterochromaffin cells. Quantitative evaluation of the immunoreaction revealed that most of the 5-HT-IR in the cytoplasm of EC cells (60%) was located over the dense cores of the secretory granules. However, a significant part of the cytoplasmic 5-HT-IR (40%) was located outside the dense cores of the secretory granules which suggests that different forms of 5-HT storage may exist.
Similar content being viewed by others
References
Ahlman H, Dahlström A (1983) Vagal mechanisms controlling serotonin release from the gastrointestinal tract and pyloric motorfunction. J Auton Nerv Syst 9:119–140
Ahlman H, Grönstad K-O, Nilsson O, Dahlström A (1984) Biochemical and morphological studies on the secretion of 5-HT into the gut lumen of the rat. Biogenic Amines 1:63–73
Barter R, Pearse AGE (1953) Detection of 5-hydroxytryptamine in mammalian enterochromaffin cells. Nature 172:810
Barter R, Pearse AGE (1955) Mammalian enterochromaffin cells as the source of serotonin (5-hydroxytryptamine). J Pathol Bacteriol 69:2531
Bendayan M, Zollinger M (1983) Ultrastructural localization of antigenic sites on osmium-fixed tissues applying the protein A-gold technique. J Histochem Cytochem 31:101–109
Dey RD, Hoffpauir J (1984) Ultrastructural immunocytochemical localization of 5-hydroxytryptamine in gastric enterochromaffin cells. J Histochem Cytochem 32:661–666
Erspamer V, Asero B (1952) Identification of enteramine, the specific hormone of the enterochromaffin cell system, as 5-hydroxytryptamine. Nature 169:800–801
Facer P, Polak JM, Jaffe BM, Pearse AGE (1979) Immunocytochemical demonstration of 5-hydroxytryptamine in gastrointestinal endocrine cells. Histochem J 11:117–121
Frens G (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature (Phys Sci) 241:20–22
Inokuchi H, Kawai K, Takeuchi Y, Sano Y (1982) Immunohistochemical demonstration of EC cells in the rat gastrointestinal tract. Histochemistry 74:453–456
Inokuchi H, Kawai K, Takeuchi Y, Sano Y (1983) Identification of EC cells in the human intestine: A comparative study between immunohistochemical and silver impregnation techniques. Histochemistry 79:9–16
Inokuchi H, Kawai K, Takeuchi Y, Sano Y (1984) Immunohistochemical study on the morphology of enterochromaffin cells in the human fundic mucosa. Cell Tissue Res 235:703–705
Lauder JM, Petrusz P, Wallace JA, Dionne A, Wilkie MB, and McCarthy K (1982) Combined serotonin immunocytochemistry and 3H-Thymidine autoradiography. J Histochem Cytochem 30:788–793
Nemoto N, Kawaoi A, Okano T, Ushiyama H, Satoh H, Shikata T (1983) Immunohistochemistry of serotonin (5-HT) in the human gut endocrine cells. An application of ultrastructural immunohistochemistry. Acta Histochem Cytochem 16:577–587
Penttilä A (1969) Identification of enterochromaffin cells in adjacent epon-embedded sections at light and electron microscopic levels. Z Zellforsch 102:193–204
Polak JM, Facer P, Pearse AGE, Jaffe BM (1978) Amine and peptide in the same APUD cell demonstrated by immunocytochemistry. Scand J Gastroenterol 13 (Suppl 49):147
Portela-Gomes GM (1982) Enterochromaffin cells, a qualitative and quantitative study. Acta Univ Ups (Suppl) 434:1982
Roth J (1982) The protein A-gold technique — a qualitative and quantitative approach for antigen localization on thin sections. In: Bullock GR, Petrusz P (eds) Techniques in immunocytochemistry, vol 1. Academic Press, New York, pp 108–133
Roth J, Bendayan M, Orci L (1978) Ultrastructural localization of intracellular antigens by the use of protein A-gold complex. J Histochem Cytochem 26:1074–1081
Schipper J, Tilders FJH (1983) A new technique for studying specificity of immunocytochemical procedures: specificity of serotonin immunostaining. J Histochem Cytochem 31:12–18
Solcia E, Capella C, Buffa R, Frigerio B (1976) Histochemical and ultrastructural studies on the argentaffin and argyrophil cells of the gut. In: Coupland RE, Fujita T (eds) Chromaffin, enterochromaffin and related cells. Elsevier Scientific Publishing Company, Amsterdam, Oxford, New York, pp 209–225
Solcia E, Polak JM, Pearse AGE, Forssman WG, Larsson L-I, Sundler F, Lechago J, Grimelius L, Fujita T, Creutzfeldt W, Gepts W, Falkmer S, Lefranc G, Heitz PH, Hage E, Buchan AMJ, Bloom SR, Grossman MI (1978) Lausanne 1977 classification of gastroenteropancreatic endocrine cells. In: Bloom SR (ed) Gut hormones. Churchill-Livingstone, Edinburgh, pp 40–48
Steinbusch HWM, Verhofstad AAJ, Joosten HWJ (1978) Localization of serotonin in the central nervous system by immunohistochemistry: description of a specific and sensitive technique and some applications. Neuroscience 3:811–819
Tange A, Takeuchi Y, Kimura H, Sano Y (1982) Immunohistochemical studies on serotonin containing cells in the gastric and duodenal mucosa of the dog. Okajimas Folia Anat Jpn 58 (Nos 4–6), pp 945–956
Vialli M, Prenna G (1969) Contribution to the cytospectrofluorimetric measurement of 5-hydroxytryptamine in enterochromaffin cells. J Histochem Cytochem 15:321–330
Wingren U, Enerbäck L, Ahlman K, Allemark S, Dahlström A (1983) Amines of the mucosal mast cell of the gut in normal and nematode infected rats. Histochemistry 77:145–158
Author information
Authors and Affiliations
Additional information
Supported by grants from the Swedish Medical Research Council (537, 2207, 5220). Göteborgs Läkaresällskap, and The Medical Faculty of Göteborg
Rights and permissions
About this article
Cite this article
Nilsson, O., Ericson, L.E., Dahlström, A. et al. Subcellular localization of serotonin immunoreactivity in rat enterochromaffin cells. Histochemistry 82, 351–355 (1985). https://doi.org/10.1007/BF00494064
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00494064