Immunoelectron microscopic and immunofluorescent localization of cytoskeletal and muscle-like contractile proteins in inner ear sensory hair cells

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Abstract

The distribution of actin, alpha-actinin, fimbrin, tropomyosin and tubulin in the apical region of inner and outer hair cells was studied by immunofluorescent localization of antibodies to these proteins. The macromolecular distribution of actin and alpha-actinin was studied using post-embedding immunoelectron microscopic techniques. Actin is present in the stereocilia and cuticular plate of both inner and outer hair cells. Antibodies to actin were localized with fluorescence and colloidal gold. Colloidal gold particles were distributed uniformly over the stereocilia, stereocilia rootlets and cuticular plate. Fimbrin is present in the stereocilia and the cuticular plate. Immunofluorescent label was more intense over the cuticular plate of outer hair cells than over the cuticular plate of inner hair cells. Alpha-actinin is present in the cuticular plate only. At the ultrastructural level, antibodies to alpha-actinin were labeled throughout the cuticular plate, with larger accumulations of colloidal gold over the electron dense bodies in the cuticular plate, as well as over the electron dense region at the junctional complex. There was no label over the electron dense portion of the stereocilia rootlets. Tropomyosin is observed in the area of the stereocilia rootlets by immunofluorescent techniques, but like fimbrin, the antigenic sites of tropomyosin did not withstand processing for ultrastructural localization. Tubulin is not present in the apical region of inner or outer hair cells, although its presence could be documented in the hair cell body and in the supporting cells.

References (48)

  • D.A. Begg et al.

    The visualization of actin filament polarity in thin sections

    J. Cell Biol.

    (1978)
  • A. Bretscher

    Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro

  • A. Bretscher et al.

    Localization of actin and microfilament-associated proteins in the microvilli and terminal web of the intestinal brush border by immunofluorescence microscopy

    J. Cell Biol.

    (1978)
  • A. Bretscher et al.

    Fimbrin, a new microfilament-associated protein present in microvilli and other cell surface structures

    J. Cell Biol.

    (1980)
  • M.C. Brown et al.

    Efferent control of cochlear inner hair cell responses in the guinea pig

    J. Physiol.

    (1984)
  • D.R. Burgess et al.

    A re-evaluation of brush border motility. Calcium induces core filament solation and microvillar vesiculation

    J. Cell Biol.

    (1982)
  • D. Drenckhahn et al.

    Localization of myosin, actin, and tropomyosin in rat intestinal epithelium: Immunohistochemical studies at the light and electron microscope levels

    J. Cell Biol.

    (1980)
  • D. Drenckhahn et al.

    Absence of myosin-like immunoreactivity in stereocilia of cochlear hair cells

    Nature (London)

    (1982)
  • D. Drenckhahn et al.

    Myosin, actin and associated proteins in the vertebrate auditory and vestibular organs. Immunocytochemical and immuno-blotting studies

  • H. Engstrom et al.

    Structure and function of the sensory hairs of the inner ear

    J. Acoust. Soc. Am.

    (1962)
  • A. Flock

    Hair cells, receptors with a motor capacity?

  • A. Flock et al.

    Actin filaments in sensory hairs of inner ear receptor cells

    J. Cell Biol.

    (1977)
  • A. Flock et al.

    Micromechanical properties of sensory hairs in receptor cells of the inner ear

    Hearing Res.

    (1983)
  • A. Flock et al.

    Studies on hair cells in isolated coils from the guinea pig cochlea

    Hearing Res.

    (1984)
  • A. Flock et al.

    Studies on the sensory hairs of receptor cells in the inner ear

    Acta Otolaryngol.

    (1977)
  • A. Flock et al.

    Immunofluorescence localization of proteins in semithin 2 μm frozen sections of the ear

    Arch. Otolaryngol.

    (1981)
  • A. Flock et al.

    Three sets of actin filaments in sensory cells of the inner ear. Identification and functional orientation determined by gel electrophoresis, immunofluorescence and electron microscopy

    J. Neurocytol.

    (1981)
  • A. Flock et al.

    Immunohistochemical localization of several cytoskeletal proteins in inner ear sensory and supporting cells

    Hearing Res.

    (1982)
  • J.R. Glenney et al.

    F-actin binding and bundling properties of fimbrin, a major cytoskeletal protein of microvillus core filaments

    J. Biol. Chem.

    (1981)
  • W.E. Gordon

    Immunofluorescent and ultrastructural studies of ‘sarcomeric’ units in stress fibers of cultured non-muscle cells

    Exp. Cell Res.

    (1978)
  • N. Hirokawa et al.

    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.

    (1982)
  • I.M. Hunter-Duvar et al.

    Anatomical changes in the organ of Corti after acoustic stimulation

  • E. Korn

    Actin polymerization and its regulation by proteins from non-muscle cells

    Physiol. Rev.

    (1982)
  • E. Lazarides

    Two general classes of cytoplasmic actin filaments in tissue culture cells; the role of tropomyosin

    J. Supramol. Struct.

    (1976)

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