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Correlative immuno-electron-microscopic and immunofluorescent localization of actin in sensory and supporting cells of the inner ear by use of a low-temperature embedding resin

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The cochleas from chinchilla inner ears were processed in the cold through Lowicryl K4M, and cured by UV light. Thick (2 μm) sections were reacted with primary antibodies raised against actin, and anti-actin antibodies localized by FITC epifluorescence. On thin sections from the same blocks anti-actin antibodies were localized ultrastructurally with secondary antibodies coupled to colloidal gold.

In the hair cells, actin was present in the stereocilia and cuticular plate, regions where thin filaments were observed by electron microscopy. Colloidal gold was uniformly distributed over these regions and over the stereocilia rootlets demonstrating that actin was present in this region although previously in permeabilized cells, the rootlet was not decorated with myosin subfragment S-1. Actin was present in the pillar and Deiters supporting cells at the reticular lamina and at the basilar membrane, where a meshwork of thin filaments was seen by electron microscopy. Colloidal gold particles were also localized over the thin processes of the pillar and Deiters cells, and over the region of the Deiters cell which envelops the base of the outer hair cell. In these regions actin co-localized with microtubules along the entire length of the supporting cells.

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  1. Angelborg C, Engstrom H (1972) Supporting elements in the organ of Corti. Acta Otolaryngol [Suppl] 301:49–60

  2. Armbruster BL, Garavito RM, Kellenberger E (1983) Dehydration and enbedding temperatures effect the antigenic specificity of tubulin and immunolabeling by the protein A-colloidal gold technique. J Histochem Cytochem 31:1380–1390

  3. Brownell WE, Bader CR, Bertrand D, de Ribaupierre Y (1985) Evoked mechanical responses of isolated cochlear outer hair cells. Science 227:194–196

  4. Drenckhahn D, Keller J, Mannherz HG, Gröschel-Stewart U, Kendrick-Jones J, Scholey J (1982) Absence of myosin-like immunoreactivity in stereocilia of cochlear hair cells. Nature 300:531–532

  5. Drenckhahn D, Prinz M, Schafer T (1984) Myosin, actin and associated proteins in the vertebrate auditory and vestibular organs. Immunocytochemical and immunoblotting studies. Abstracts of the Midwinter Meeting, Assoc Res Otolaryngol 7:139–140

  6. Flock A (1983) Hair cells, receptors with a motor capacity? In: Klink R, Hartman R (eds) Hearing-physiological bases and psychophysics. Springer, Berlin Heidelberg New York Tokyo, PP 1–6

  7. Flock A, Cheung HC (1977) Actin filaments in sensory hairs of inner ear receptor cells. J Cell Biol 75:339–343

  8. Flock A, Cheung HC, Flock B, Utter G (1981) Three sets of actin filaments in sensory cells of the inner ear. J Neurocytol 10:133–147

  9. Flock A, Bretscher A, Weber K (1982) Immunohistochemical localization of several cytoskeletal proteins in inner ear sensory and supporting cells. Hear Res 6:75–89

  10. Giloh H, Sedat JW (1982) Fluorescence microscopy: reduced photobleaching of rhodamine and fluorescein protein conjugates by n-propyl gallate. Science 217:1252–1255

  11. Hirokawa N, Tilney L (1982) Interactions between actin filaments and between actin filaments and membrane in quick-frozen and deeply etched hair cells of the chick ear. J Cell Biol 95:249–261

  12. Hudspeth AJ, Jacobs R (1979) Stereocilia mediate transduction in vertebrate hair cells. Proc Natl Acad Sci USA 76:1506–1509

  13. Hunter-Duvar IM, Suzuki M, Mount RJ (1982) Anatomical changes in the organ of Corti after acoustic stimulation. In: Hamernik RP, Henderson D, Salvi R (eds) New perspectives on noise induced hearing loss. Raven Press, New York, pp 3–21

  14. Lemanski LF, Paulson DJ, Hill CS, Davis LA, Riles LC, Lim SS (1985) Immunoelectron microscopic localization of alpha-actinin on thin-sectioned tissues. J Histochem Cytochem 33:515–522

  15. Macartney JC, Comis SD, Pickles JP (1980) Is myosin in the cochlea a basis for active motility? Nature 288:491–492

  16. McLean IW, Nakane PK (1974) Periodate-lysine-paraformaldehyde fixative, a new fixative for immunoelectron microscopy. J Histochem Cytochem 22:1077–1086

  17. Maupin-Szamier P, Pollard TD (1978) Actin filament destruction by osmium tetroxide. J Cell Biol 77:837–852

  18. Orman S, Flock A (1983) Active control of sensory hair mechanics implied by susceptibility to media that induce contraction in muscle. Hear Res 11:261–266

  19. Roth J, Bendayan M, Carlemalm E, Villiger W, Garavito M (1981) Enhancement of structural preservation and immunocytochemical staining in low temperature embedded pancreatic tissue. J Histochem Cytochem 29:663–671

  20. Slepecky N, Hamernik RP, Henderson D (1980) A re-examination of a hair cell organelle in the cuticular plate region and its possible relation to active processes in the cochlea. Hear Res 2:413–421

  21. Slepecky N, Chamberlain SC (1982) Distribution and polarity of actin in the sensory hair cells of the chinchilla cochlea. Cell Tissue Res 224:15–24

  22. Slepecky N, Chamberlain SC (1983) Distribution and polarity of actin in inner ear supporting cells. Hear Res 10:359–370

  23. Tilney LG, De Rosier DJ, Mulroy MJ (1980) The organization of actin filaments in the stereocilia of cochlear hair cells. J Cell Biol 86:244–259

  24. Zenner HP (1981) Cytoskeletal and muscle-like elements in cochlear hair cells. Arch Otorhinolaryngol 230:81–92

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Correspondence to Dr. Norma Slepecky.

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Slepecky, N., Chamberlain, S.C. Correlative immuno-electron-microscopic and immunofluorescent localization of actin in sensory and supporting cells of the inner ear by use of a low-temperature embedding resin. Cell Tissue Res. 245, 229–235 (1986).

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Key words

  • Inner ear
  • Cochlea
  • Actin
  • Immunocytochemistry
  • Chinchilla