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Ultrastructural colocalization of the bioactive mediators 5-hydroxytryptamine and bombesin in endocrine cells of human fetal airways

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Summary

Endocrine cells in the airway epithelium of human fetal lungs are known to contain an amine, 5-hydroxytryptamine (5HT), and a peptide, bombesin (BOM). These mediators may be involved in regulating smooth muscle and secretory activity in the airways as well as in development of the fetal lung. However, the exact endocrine cell type that contains 5HT and BOM has not been described at the ultrastructural level. This investigation provides immunocytochemical evidence that 5HT and BOM are stored in a single cell type, the P1 cell. Thin sections of airways from human fetal lungs were incubated either in anti-5HT antiserum (diluted 1∶3000) or in anti-BOM antiserum (diluted 1∶600) and then labeled with affinity purified goat anti-rabbit IgG coupled to 16 nm gold particles. For colocalization, thin sections were incubated on one side to demonstrate 5HT and on the other side to demonstrate BOM. Two different sizes of gold particles (10 and 30 nm) were coupled to IgGs and used for the labeled second antibodies. Controls consisted of absorbing of the primary antiserum with an excess of either 5HT or BOM. 5HT-and BOM-like immunoreactivities were observed in the dense-core vesicles (DCV) of P1 cells, and it was apparent from serial sections that 5HT and BOM labeling was sometimes present in the same P1 cells. Sections labeled for 5HT on one side with large gold particles and for BOM on the other side with small gold particles revealed that 5HT-and BOM-like immunoreactivities were located in the same DCV. Labeling did not occur when the anti-5HT antiserum was absorbed with 5HT or when the anti-BOM antiserum was absorbed with BOM. These results demonstrate that 5HT-and BOM-like immunoreactivity is present in P1 endocrine cells of human fetal lung. Furthermore, a single DCV contains both 5HT and BOM.

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References

  • Becker KL, Monaghan KG, Silva OL (1980) Immunocytochemical localization of calcitonin in Kultchitsky cells of human lung. Arch Pathol Lab Med 104:196–198

    Google Scholar 

  • Bendayan M, Stephens H (1984) Double labeling cytochemistry applying the protein A-gold technique. In: Polak JM, Varndell IM (eds) Immunolabelling for electron microscopy. Elsevier, Amsterdam, pp 143–154

    Google Scholar 

  • Cutz E, Chan W, Track NS (1981) Bombesin, calcitonin and leuenkephalin immunoreactivity in endocrine cells of human lung. Experimentia 37:765–767

    Google Scholar 

  • Cutz E, Chan W, Wong V, Conen PE (1975) Ultrastructure and fluorescence histochemistry of endocrine (APUD-type) cells in tracheal mucosa of human and various animal species. Cell Tissue Res 158:425–437

    Google Scholar 

  • Cutz E, Gillan JE, Track NS (1984) Pulmonary endocrine cells in the developing human lung and during neonatal adaptation. In: Becker KL, Gazdar AF (eds) The endocrine lung in health and disease. WB Saunders, Philadelphia, pp 210–231

    Google Scholar 

  • Ericson LE, Hakanson R, Larsson B, Owman Ch, Sundler F (1972) Fluorescence and electron microscopy of amine-storing enterochromaffin-like cells in the tracheal epithelium of mouse. Z Zellforsch 124:532–545

    Google Scholar 

  • Erisman MD, Linnoila RI, Hernandez O, DiAugustine RP, Lazarus LF (1982) Human lung small-cell carcinoma contains bombesin. Proc Natl Acad Sci USA 79:2379–2383

    Google Scholar 

  • Feyrter F (1953) Über die peripheren endokrinen (parakrinen) Drüsen des Menschen. Wien Düsseldorf, W. Maudrich

    Google Scholar 

  • Frens G (1973) Controlled nucleation for the regulation of particle size in monodisperse gold suspensions. Nature Phys Sci 241:20–22

    Google Scholar 

  • Fröhlich F (1949) Die „Hellen Zellen” der Bronchialschleimhaut und ihre Beziehungen zum Problem der Chemoreceptoren. Frankfurter Z Pathol 60:517–559

    Google Scholar 

  • Ghaeti MA, Sheppard MN, Henzen-Logman S, Blank MA, Polak JM, Bloom SR (1983) Bombesin and vasoactive intestinal polypeptide in the developing lung: Marked changes in acute respiratory distress syndrome. J Clin Endocrinol Metab 57:1226–1232

    Google Scholar 

  • Hage E (1971) Endocrine cells in the bronchial mucosa of human fetuses. Acta Pathol Microbiol Scand [Section A] 79:307–308

    Google Scholar 

  • Hage E (1973) Amine-handling properties of APUD-cells in the bronchial epithelium of human fetuses and in the epithelium of the main bronchi of human adults. Acta Pathol Microbiol Scand [Section A] 81:64–70

    Google Scholar 

  • Hage E (1980) Light and electron microscopic characteristics of the various lung endocrine cell types. Invest Cell Pathol 3:345–351

    Google Scholar 

  • Hernandez-Vasquez A, Will JA, Quay WB (1977) Quantitative characteristics of the Feyrter (APUD) cells of the neonatal rabbit lung in normoxia and chronic hypoxia. Thorax 32:449–456

    Google Scholar 

  • Hökfelt T, Johansson O, Goldstein M (1984) Chemical anatomy of the brain. Science 225:1326–1334

    Google Scholar 

  • Hökfelt T, Johansson O, Ljungdahl A, Lundberg JM, Schultzberg M (1980) Peptidergic neurones. Nature 284:515–521

    Google Scholar 

  • Impicciatore M, Bertaccini G (1973) The bronchoconstrictor action of the tetradecapeptide bombesin in the guinea pig. J Pharm Pharmacol 25:872–875

    Google Scholar 

  • Johnson RG, Beers MF, Scarpa A (1982a) H+ ATPase of chromaffin granules. J Biol Chem 257:10701–10707

    Google Scholar 

  • Johnson DE, Lock JE, Elde RP, Thompson TR (1982b) Pulmonary neuroendocrine cells in hyaline membrane disease and bronchopulmonary displasia. Pediatr Res 16:446–454

    Google Scholar 

  • Kobayashi S (1975) Uptake and intracellular localization of exogenous L-DOPA, L-lecuine and their metabolites in the gastroenteric endocrine cells of the mouse studied by electron microscope autoradiography. Acta Histol Jpn 37:313–333

    Google Scholar 

  • Kolata G (1982) Cell biology yields clues to lung cancer. Science 218:38–39

    Google Scholar 

  • Kulik TJ, Johnson DE, Elde RP, Locke JE (1983) Pulmonary vascular effects of bombesin and gastrin-releasing peptide in conscious newborn lambs. J Appl Physiol 55:1093–1097

    Google Scholar 

  • Larsson L-I (1980) On the possible existence of multiple endocrine, paracrine, and neurocrine messenges in secretory cell systems. Invest Cell Pathol 3:73–85

    Google Scholar 

  • Lauweryns JM, Cokelaere M (1973) Hypoxia-sensitive neuroepithelial bodies: intrapulmonary secretory neuroreceptors, modulated by the CNS. Z Zellforsch 145:521–540

    Google Scholar 

  • Lauweryns JM, Peuskens JC, Cokelaere M (1970) Argyrophil, fluorescent and granulated (peptide and amine producing?) AFG cells in human infant bronchial epithelium. Life Sci 9:1417–1429

    Google Scholar 

  • Lehy T, Accary JP, LaBeille D, Dubrasquet M (1983) Chronic administration of bombesin stimulates antral gastrin cell proliferation in the rat. Gastroenterology 84:914–919

    Google Scholar 

  • Lundberg JM, Anggard A, Fahrenkrug J, Hökfelt T, Mutt V (1980) Vasoactive intestinal polypeptide in cholinergic neurons of exocrine glands: functional significance of coexisting transmitters for vasodilation and secretion. Proc Natl Acad Sci USA 77:1651–1655

    Google Scholar 

  • Marchevsky AM, Kleinerman J (1983) Quantitative and immunocytochemical studies of APUD cells in airway and gut. Lung 161:67–76

    Google Scholar 

  • Moody TW, Pert CB, Gazdar AF, Carney DN, Minna JD (1981) High levels of intracellular bombesin characterize human smallcell carcinoma. Science 214:1246–1248

    Google Scholar 

  • Mühlpfordt H (1982) The preparation of colloidal gold particles using tannic acid as an additional reducing agent. Experientia 38:1127–1128

    Google Scholar 

  • Near JA, Mahler HR (1983) Reserpine labels the catecholamine transporter in synaptic vesicles from bovine caudate nucleus. FEBS Lett 158:31–35

    Google Scholar 

  • Nunez EA, Gershon MD (1972) Synthesis and storage of serotonin by parafollicular (C) cells of the thyroid gland of active, prehibernating and hibernating bats. Endocrinology 90:1008–1024

    Google Scholar 

  • Pearse AGE, Takor Takor T (1979) Embryology of the diffuse neuroendocrine system and its relationship to the common peptides. Fed Proc 38:2288–2294

    Google Scholar 

  • Pelletier G, Steinbusch HWM, Verhofstad AAJ (1981) Immunoreactive substance P and serotonin present in the same densecore vesicles. Nature 293:71–72

    Google Scholar 

  • Price J, Penman E, Bourne GL, Rees LH (1983) Characterization of bombesin-like immunoreactivity in human fetal lung. Regul Pept 7:315–322

    Google Scholar 

  • 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

    Google Scholar 

  • Rozengurt E, Sinnett-Smith J (1983) Bombesin stimulation of DNA synthesis and cell division in cultures of Swiss 3T3 cells. Proc Natl Acad Sci USA 80:2936–2940

    Google Scholar 

  • Said SI, Mutt V, Erdos EG (1980) The lung in relation to vasoactive polypeptides. Ciba Foundation Symposium 78:217–237

    Google Scholar 

  • Stahlman MT, Gray ME (1984) Ontogony of neuroendocrine cells in human fetal lung. I. An electron microscopic study. Lab Invest 51:449–463

    Google Scholar 

  • Stahlman MT, Kasselberg AG, Orth DN, Gray ME (1985) Ontogeny of neuroendocrine cells in human fetal lung. II. An immunohistochemical study. Lab Invest 52:52–60

    Google Scholar 

  • Track NS, Cutz E (1982) Bombesin-like immunoreactivity in developing human lung. Life Sci 30:1553–1556

    Google Scholar 

  • Tsutsumi Y, Osamura RY, Wantanabe K, Yanaihara N (1983) Simultaneous immunohistochemical localization of gastrin releasing peptide (GRP) and calcitonin (CT) in human bronchial endocrine-type cells. Virchows Arch [Pathol Anat] 400:163–171

    Google Scholar 

  • Wharton J, Polak JM, Bloom SR, Ghatei MA, Solcia E, Brown MR, Pearse AGE (1978) Bombesin-like immunoreactivity in the lung. Nature 273:769–770

    Google Scholar 

  • Wharton J, Polak JM, Cole GA, Marangos PJ, Pearse AGE (1981) Neuron-specific enolase as an immunocytochemical marker for the diffuse neuroendocrine system in human fetal lung. J Histochem Cytochem 29:1359–1364

    Google Scholar 

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  1. Department of Anatomy, West Virginia University Medical Center, Morgantown, West Virginia, USA

    Richard D. Dey Ph.D. & John M. Hoffpauir

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  1. Richard D. Dey Ph.D.
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  2. John M. Hoffpauir
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Dey, R.D., Hoffpauir, J.M. Ultrastructural colocalization of the bioactive mediators 5-hydroxytryptamine and bombesin in endocrine cells of human fetal airways. Cell Tissue Res. 246, 119–124 (1986). https://doi.org/10.1007/BF00219007

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