Abstract
The immunohistochemical characteristics of brush cells in the laryngeal mucosa were examined using immunohistochemistry for various immunohistochemical cell markers including villin at the light and electron microscopic levels. Cells that were immunoreactive to villin were barrel-shaped with thick cytoplasmic processes extending toward the lumen of the laryngeal cavity. Immunoelectron microscopic observations revealed thick and short microvilli with long rootlets of microfilaments. Numerous small clear vesicles and small finger-like cytoplasmic processes were observed in the apical process and lateral membrane, respectively. Double immunofluorescence showed villin-immunoreactive cells were not immunoreactive for the markers of solitary chemosensory cells, GNAT3 and phospholipase C, β2-subunit (PLCβ2), or for that of neuroendocrine cells, synaptosome-associated protein 25kD. Furthermore, immunoreactivities for cytokeratin 18 (CK18) and doublecortin like-kinase 1 in the perinuclear cytoplasm of villin-immunoreactive cells. However, some CK18-immunoreactive cells were immunoreactive to GNAT3 but not to villin. Regarding sensory innervation, only a few intraepithelial nerve endings with P2X3, SP, or CGRP immunoreactivity attached to villin-immunoreactive cells. In the present study, brush cells in the rat laryngeal mucosa were classified by immunoreactivity for villin, and were independent of other non-ciliated epithelial cells such as solitary chemosensory cells and neuroendocrine cells.
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References
Bezençon C, Fürholz A, Raymond F et al (2008) Murine intestinal cells expressing Trpm5 are mostly brush cells and express markers of neuronal and inflammatory cells. J Comp Neurol. 509:514–525
Bjerknes M, Khandanpour C, Möröy T et al (2012) Origin of the brush cell lineage in the mouse intestinal epithelium. Dev Biol 362:194–218. https://doi.org/10.1016/j.ydbio.2011.12.009
Deckmann K, Kummer W (2016) Chemosensory epithelial cells in the urethra: sentinels of the urinary tract. Histochem Cell Biol 146:673–683. https://doi.org/10.1007/s00418-016-1504-x
Finger TE (2005) ATP Signaling is crucial for communication from taste buds to gustatory nerves. Science 310:1495–1499. https://doi.org/10.1126/science.1118435
Gerbe F, Brulin B, Makrini L et al (2009) DCAMKL-1 expression identifies Tuft cells rather than stem cells in the adult mouse intestinal epithelium. Gastroenterology 137:2179–2180. https://doi.org/10.1053/j.gastro.2009.06.072
Gerbe F, van Es JH, Makrini L et al (2011) Distinct ATOH1 and Neurog3 requirements define tuft cells as a new secretory cell type in the intestinal epithelium. J Cell Biol 192:767–780. https://doi.org/10.1083/jcb.201010127
Hass N, Schwarzenbacher K, Breer H (2007) A cluster of gustducin-expressing cells in the mouse stomach associated with two distinct populations of enteroendocrine cells. Histochem Cell Biol 128:457–471. https://doi.org/10.1007/s00418-007-0325-3
Hijiya K, Okada Y, Tankawa H (1977) Ultrastructural study of the alveolar brush cell. J Electron Microsc 26:321–329. https://doi.org/10.1093/oxfordjournals.jmicro.a050078
Höfer D, Drenckhahn D (1992) Identification of brush cells in the alimentary and respiratory system by antibodies to villin and fimbrin. Histochemistry 98:237–242
Höfer D, Drenckhahn D (1996) Cytoskeletal markers allowing discrimination between brush cells and other epithelial cells of the gut including enteroendocrine cells. Histochem Cell Biol 105:405–412
Höfer D, Drenckhahn D (1998) Identification of the taste cell G-protein, α-gustducin, in brush cells of the rat pancreatic duct system. Histochem Cell Biol 110:303–309
Höfer D, Püschel B, Drenckhahn D (1996) Taste receptor-like cells in the rat gut identified by expression of α-gustducin. Proc Natl Acad Sci USA 93:6631–6634
Hoover B, Baena V, Kaelberer MM et al (2017) The intestinal tuft cell nanostructure in 3D. Sci Rep 7:1652. https://doi.org/10.1038/s41598-017-01520-x
Jeffery PK, Reid L (1975) New observations of rat airway epithelium: a quantitative and electron microscopic study. J Anat 120:295–320
Kaske S, Krasteva G, König P et al (2007) TRPM5, a taste-signaling transient receptor potential ion-channel, is a ubiquitous signaling component in chemosensory cells. BMC Neurosci 8:49. https://doi.org/10.1186/1471-2202-8-49
Kasper M, Höfer D, Woodcock-Mitchell J et al (1994) Colocalization of cytokeratin 18 and villin in type III alveolar cells (brush cells) of the rat lung. Histochemistry 101:57–62
Krasteva G, Canning BJ, Hartmann P et al (2011) Cholinergic chemosensory cells in the trachea regulate breathing. Proc Natl Acad Sci USA 108:9478–9483. https://doi.org/10.1073/pnas.1019418108
Lewis DJ, Prentice DE (1980) The ultrastructure of rat laryngeal epithelia. J Anat 130:617–632
Luciano L, Reale E, Ruska H (1968) Über eine “chemorezeptive” Sinneszelle in der Trachea der Ratte. Z Zellforsch Mikrosk Anat 85:350–375. https://doi.org/10.1007/BF00328847
Luciano L, Reale E, Ruska H (1969) Bürstenzellen im Alveolarepithel der Rattenlunge. Z Zellforsch Mikrosk Anat 95:198–201. https://doi.org/10.1007/BF00968452
Marin ML, Lane BP, Gordon RE, Drummond E (1979) Ultrastructure of rat tracheal epithelium. Lung 156:223–236
McDowell EM, Sorokin SP, Hoyt RF (1994) Ontogeny of endocrine cells in the respiratory system of Syrian golden hamsters. I. Larynx and trachea. Cell Tissue Res 275:143–156
Meyrick B, Reid L (1968) The alveolar brush cell in rat lung - a third pneumonocyte. J Ultrastruct Res 23:71–80
Reid L, Meyrick B, Antony VB et al (2005) The mysterious pulmonary brush cell: a cell in search of a function. Am J Respir Crit Care Med 172:136–139
Saqui-Salces M, Keeley TM, Grosse AS et al (2011) Gastric tuft cells express DCLK1 and are expanded in hyperplasia. Histochem Cell Biol 136:191–204. https://doi.org/10.1007/s00418-011-0831-1
Sato A (2007) Tuft cells. Anat Sci Int 82:187–199. https://doi.org/10.1111/j.1447-073X.2007.00188.x
Sato A, Miyoshi S (1997) Fine structure of tuft cells of the main excretory duct epithelium in the rat submandibular gland. Anat Rec 248:325–331
Saunders CJ, Reynolds SD, Finger TE (2013) Chemosensory brush cells of the trachea. A stable population in a dynamic epithelium. Am J Respir Cell Mol Biol 49:190–196. https://doi.org/10.1165/rcmb.2012-0485OC
Sbarbati A, Osculati F (2005a) A new fate for old cells: brush cells and related elements. J Anat 206:349–358. https://doi.org/10.1111/j.1469-7580.2005.00403.x
Sbarbati A, Osculati F (2005b) The taste cell-related diffuse chemosensory system. Prog Neurobiol 75:295–307. https://doi.org/10.1016/j.pneurobio.2005.03.001
Sekerková G, Zheng L, Loomis PA et al (2006) Espins and the actin cytoskeleton of hair cell stereocilia and sensory cell microvilli. Cell Mol Life Sci 63:2329–2341. https://doi.org/10.1007/s00018-006-6148-x
Souma T (1987) The distribution and surface ultrastructure of airway epithelial cells in the rat lung: a scanning electron microscopic study. Arch Histol Jpn 50:419–436
Taira K, Shibasaki S (1978) A fine structure study of the non-ciliated cells in the mouse tracheal epithelium with special reference to the relation of “brush cells” and “endocrine cells”. Arch Histol Jpn 41:351–366. https://doi.org/10.1679/aohc1950.41.351
Takahashi N, Nakamuta N, Yamamoto Y (2016) Morphology of P2 × 3-immunoreactive nerve endings in the rat laryngeal mucosa. Histochem Cell Biol 145:131–146. https://doi.org/10.1007/s00418-015-1371-x
Tizzano M, Cristofoletti M, Sbarbati A, Finger TE (2011) Expression of taste receptors in solitary chemosensory cells of rodent airways. BMC Pulm Med 11:3. https://doi.org/10.1186/1471-2466-11-3
Yamamoto Y, Kusakabe T, Hayashida Y et al (2000) Laryngeal endocrine cells: topographic distribution and adaptation to chronic hypercapnic hypoxia. Histochem Cell Biol 114:277–282
Yu YC, Miyazaki J, Shin T (1996) Neuroendocrine cells in the cat laryngeal epithelium. Eur Arch Otorhinolaryngol 253:287–293
Acknowledgements
The authors are grateful to Ms. Eri Ishiyama, Technical Support Center for Life Science Research, Iwate Medical University for her technical support with the EM study. This work was supported by the JSPS KAKENHI: Grant Number 15K07759 (YY) from the Japan Society for the Promotion of Science (JSPS), Japan.
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Yamamoto, Y., Ozawa, Y., Yokoyama, T. et al. Immunohistochemical characterization of brush cells in the rat larynx. J Mol Hist 49, 63–73 (2018). https://doi.org/10.1007/s10735-017-9747-y
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DOI: https://doi.org/10.1007/s10735-017-9747-y