Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1989 Oct 1;109(4):1711–1723. doi: 10.1083/jcb.109.4.1711

Preliminary biochemical characterization of the stereocilia and cuticular plate of hair cells of the chick cochlea

PMCID: PMC2115824  PMID: 2677026

Abstract

The sensory epithelium of the chick cochlea contains only two cell types, hair cells and supporting cells. We developed methods to rapidly dissect out the sensory epithelium and to prepare a detergent-extracted cytoskeleton. High salt treatment of the cytoskeleton leaves a "hair border", containing actin filament bundles of the stereocilia still attached to the cuticular plate. On SDS-PAGE stained with silver the intact epithelium is seen to contain a large number of bands, the most prominent of which are calbindin and actin. Detergent extraction solubilizes most of the proteins including calbindin. On immunoblots antibodies prepared against fimbrin from chicken intestinal epithelial cells cross react with the 57- and 65-kD bands present in the sensory epithelium and the cytoskeleton. It is probable that the 57-kD is a proteolytic fragment of the 65-kD protein. Preparations of stereocilia attached to the overlying tectorial membrane contain the 57- and 65-kD bands. A 400-kD band is present in the cuticular plate. By immunofluorescence, fimbrin is detected in stereocilia but not in the hair borders after salt extraction. The prominent 125 A transverse stripping pattern characteristic of the actin cross-bridges in a bundle is also absent in hair borders suggesting fimbrin as the component that gives rise to the transverse stripes. Because the actin filaments in the stereocilia of hair borders still remain as compact bundles, albeit very disordered, there must be an additional uncharacterized protein besides fimbrin that cross-links the actin filaments together.

Full Text

The Full Text of this article is available as a PDF (6.9 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bretscher A. Fimbrin is a cytoskeletal protein that crosslinks F-actin in vitro. Proc Natl Acad Sci U S A. 1981 Nov;78(11):6849–6853. doi: 10.1073/pnas.78.11.6849. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bretscher A., Weber K. Fimbrin, a new microfilament-associated protein present in microvilli and other cell surface structures. J Cell Biol. 1980 Jul;86(1):335–340. doi: 10.1083/jcb.86.1.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Coluccio L. M., Bretscher A. Reassociation of microvillar core proteins: making a microvillar core in vitro. J Cell Biol. 1989 Feb;108(2):495–502. doi: 10.1083/jcb.108.2.495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DeRosier D. J., Tilney L. G., Egelman E. Actin in the inner ear: the remarkable structure of the stereocilium. Nature. 1980 Sep 25;287(5780):291–296. doi: 10.1038/287291a0. [DOI] [PubMed] [Google Scholar]
  5. DeRosier D. J., Tilney L. G. How actin filaments pack into bundles. Cold Spring Harb Symp Quant Biol. 1982;46(Pt 2):525–540. doi: 10.1101/sqb.1982.046.01.049. [DOI] [PubMed] [Google Scholar]
  6. DeRosier D. J., Tilney L. G. The form and function of actin. A product of its unique design. Cell Muscle Motil. 1984;5:139–169. doi: 10.1007/978-1-4684-4592-3_3. [DOI] [PubMed] [Google Scholar]
  7. Drenckhahn D., Dermietzel R. Organization of the actin filament cytoskeleton in the intestinal brush border: a quantitative and qualitative immunoelectron microscope study. J Cell Biol. 1988 Sep;107(3):1037–1048. doi: 10.1083/jcb.107.3.1037. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Drenckhahn D., Kellner J., Mannherz H. G., Gröschel-Stewart U., Kendrick-Jones J., Scholey J. Absence of myosin-like immunoreactivity in stereocilia of cochlear hair cells. Nature. 1982 Dec 9;300(5892):531–532. doi: 10.1038/300531a0. [DOI] [PubMed] [Google Scholar]
  9. Glenney J. R., Jr, Kaulfus P., Matsudaira P., Weber K. F-actin binding and bundling properties of fimbrin, a major cytoskeletal protein of microvillus core filaments. J Biol Chem. 1981 Sep 10;256(17):9283–9288. [PubMed] [Google Scholar]
  10. Hirokawa N. The ultrastructure of the basilar papilla of the chick. J Comp Neurol. 1978 Sep 15;181(2):361–374. doi: 10.1002/cne.901810208. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Macartney J. C., Comis S. D., Pickles J. O. Is myosin in the cochlea a basis for active motility? Nature. 1980 Dec 4;288(5790):491–492. doi: 10.1038/288491a0. [DOI] [PubMed] [Google Scholar]
  13. Maupin-Szamier P., Pollard T. D. Actin filament destruction by osmium tetroxide. J Cell Biol. 1978 Jun;77(3):837–852. doi: 10.1083/jcb.77.3.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Oberholtzer J. C., Schneider M. E., Summers M. C., Saunders J. C., Matschinsky F. M. The developmental appearance of a major basilar papilla-specific protein in the chick. Hear Res. 1986;23(2):161–168. doi: 10.1016/0378-5955(86)90013-4. [DOI] [PubMed] [Google Scholar]
  15. Slepecky N., Chamberlain S. C. Immunoelectron microscopic and immunofluorescent localization of cytoskeletal and muscle-like contractile proteins in inner ear sensory hair cells. Hear Res. 1985;20(3):245–260. doi: 10.1016/0378-5955(85)90029-2. [DOI] [PubMed] [Google Scholar]
  16. Sobin A., Flock A. Immunohistochemical identification and localization of actin and fimbrin in vestibular hair cells in the normal guinea pig and in a strain of the waltzing guinea pig. Acta Otolaryngol. 1983 Nov-Dec;96(5-6):407–412. doi: 10.3109/00016488309132726. [DOI] [PubMed] [Google Scholar]
  17. Takasaka T., Smith C. A. The structure and innervation of the pigeon's basilar papilla. J Ultrastruct Res. 1971 Apr;35(1):20–65. doi: 10.1016/s0022-5320(71)80141-7. [DOI] [PubMed] [Google Scholar]
  18. Tanaka K., Smith C. A. Structure of the chicken's inner ear: SEM and TEM study. Am J Anat. 1978 Oct;153(2):251–271. doi: 10.1002/aja.1001530206. [DOI] [PubMed] [Google Scholar]
  19. Tilney L. G., DeRosier D. J. Actin filaments, stereocilia, and hair cells of the bird cochlea. IV. How the actin filaments become organized in developing stereocilia and in the cuticular plate. Dev Biol. 1986 Jul;116(1):119–129. doi: 10.1016/0012-1606(86)90048-5. [DOI] [PubMed] [Google Scholar]
  20. Tilney L. G., Egelman E. H., DeRosier D. J., Saunder J. C. Actin filaments, stereocilia, and hair cells of the bird cochlea. II. Packing of actin filaments in the stereocilia and in the cuticular plate and what happens to the organization when the stereocilia are bent. J Cell Biol. 1983 Mar;96(3):822–834. doi: 10.1083/jcb.96.3.822. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Tilney L. G., Saunders J. C. Actin filaments, stereocilia, and hair cells of the bird cochlea. I. Length, number, width, and distribution of stereocilia of each hair cell are related to the position of the hair cell on the cochlea. J Cell Biol. 1983 Mar;96(3):807–821. doi: 10.1083/jcb.96.3.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamashiro-Matsumura S., Matsumura F. Purification and characterization of an F-actin-bundling 55-kilodalton protein from HeLa cells. J Biol Chem. 1985 Apr 25;260(8):5087–5097. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES