Skip to main content
Log in

Immunolocalization of peptide 19 and other calcium-binding proteins in the guinea pig cochlea

  • Original Articles
  • Published:
Anatomy and Embryology Aims and scope Submit manuscript

Abstract

Calcium ions are known to play critical roles in a variety of cochlear functions. The distributions of a number of calcium binding proteins that regulate calcium ion levels within the cochlea have previously been described. In this report we extend and refine previous reports of the distribution of immunostaining for calmodulin, calbindin, and calretinin and show for the first time the distribution for peptide 19. There were longitudinal and radial gradients of immunostaining for peptide 19 within outer hair cells that appeared to match previously described gradients of efferent innervation of these cells. Gradients of immunostaining for calbindin within outer hair cells were in the opposite directions, which suggests that levels of this protein are correlated with afferent innervation density and perhaps the abundance of subsurface cisternae. No gradients were seen in the distributions of cells stained for calmodulin and calretinin, which included sensory cells and supporting cells respectively. All ganglion cells were stained for calmodulin but the other proteins appeared to be present in limited ganglion cell subpopulations. In addition to staining of sensorineural elements, antisera to all compounds but peptide 19 showed immunostaining of cells within the lateral wall and the spiral limbus. The results suggest that the proteins under study are involved in a wide variety of calcium-regulated functions within the cochlea. Knowledge of the unique distribution of each of the compounds should facilitate further studies of their roles in cochlear function.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Adams JC (1992) Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40: 1457–1463

    Google Scholar 

  • Andressen C, Blumcke I, Celio MR (1993) Calcium-binding proteins: selective marker of nerve cells. Cell Tissue Res 271: 181–208

    CAS  PubMed  Google Scholar 

  • Baimbridge KG, Miller JI, Parkes CO (1982) Calcium-binding protein distribution in the rat brain. Brain Res 239: 519–525

    Google Scholar 

  • Berrebi AS, Oberdick J, Sangameswaran L, Christakos S, Morgan JI, Mugnaini E (1991) Cerebellar Purkinje cell markers are expressed in retinal bipolar neurons. J Comp Neurol 308: 630–649

    Google Scholar 

  • Brown MC (1987) Morphology of labeled efferent fibers in the guinea pig cochlea. J Comp Neurol 260: 605–618

    Google Scholar 

  • Brundin L, Flock Å, Canlon B (1989) Sound-induced motility of isolated cochlea outer hair cells is frequency-specific. Nature 342: 814–816

    Google Scholar 

  • Canlon B, Brundin L, Flock Å (1988) Acoustic stimulation causes tonotopic alterations in the length of isolated outer hair cells from guinea pig hearing organ. Proc Natl Acad Sci USA 85: 7033–7035

    Google Scholar 

  • Celio MR, Scharer L, Morrison JH, Norman AW, Bloom FE (1986) Calbindin immunoreactivity alternates with cytochrome C oxidase-rich zones in some layers of primate visual cortex. Nature 323: 715–717

    Google Scholar 

  • Crawford AC, Evans MG, Fettiplace R (1991) The actions of calcium on the mechano-electrical transducer current of turtle hair cells. J Physiol 434: 369–398

    Google Scholar 

  • Dechesne CJ, Thomasset M (1988) Calbindin (CaBP 28 kDa) appearance and distribution during development of the mouse inner ear. Brain Res Dev Brain Res 40: 233–242

    Google Scholar 

  • Dechesne CJ, Lavigne-Rebillard M, Brehier A, Thomasset M, Sans A (1988a) Appearance and distribution of neuron-specific enolase and calbindin (CaBP 28 kDa) in the developing human inner ear. Brain Res Dev Brain Res 41: 221–230

    Google Scholar 

  • Dechesne CJ, Brehier A, Thomasset M, Sans A (1988b) Calbindin (CaBP 28 kDa) localization in the peripheral vestibular system of various vertebrates. Hear Res 33: 273–278

    Google Scholar 

  • Dechesne CJ, Winsky L, Kim HN, Goping G, Vu TD, Wenthold RJ, Jacobowitz DM (1991) Identification and ultrastructural localization of a calretinin-like calcium-binding protein (protein 10) in the guinea pig and rat inner ear. Brain Res: 560: 139–148

    Google Scholar 

  • Dechesne CJ, Winsky L, Moniot B, Raymond J (1993) Localization of calretinin mRNA in rat and guinea pig inner ear by in situ hybridization using radioactive and non-radioactive probes. Hear Res 69: 91–97

    Google Scholar 

  • Desmadryl G, Dechesne CJ (1992) Calretinin immunoreactivity in chinchilla and guinea pig vestibular end organs characterizes the calyx unit subpopulation. Exp Brain Res 89: 105–108

    Google Scholar 

  • Dulon D, Zajic G, Schacht J (1991) Differential motile response of isolated inner and outer hair cells to stimulation by potassium and calcium ions. Hear Res 52: 225–232

    Google Scholar 

  • Dunn RA, Morest DK (1975) Receptor synapses without synaptic ribbons in the cochlea of the cat. Proc Natl Acad Sci USA 72: 3599–3603

    Google Scholar 

  • Eatock RA, Corey DP, Hudspeth AJ (1987) Adaptation of mechanoelectrical transduction in hair cells of bullfrog's sacculus. J Neurosci 9: 2821–2836

    Google Scholar 

  • Fettiplace R (1992) The role of calcium in hair cell transduction. In: Corey DP, Roper SD (eds) Sensory transduction. Rockefeller University Press, New York, pp 333–356

    Google Scholar 

  • Flock Å, Flock B, Ulfendahl M (1986) Mechanisms of movement in outer hair cells and possible structural basis. Arch Otorhinolaryngol 243: 83–90

    CAS  PubMed  Google Scholar 

  • Furness DN, Hackney CM (1990) Comparative ultrastructure of subsurface cisternae in inner and outer hair cells of the guinea pig cochlea. Eur Arch Otorhinolaryngol 247: 12–15

    Google Scholar 

  • Gulley RL, Reese TS (1997) Freeze-fracture studies on the synapses in the organ of Corti. J Comp Neurol 171: 517–544

    Google Scholar 

  • Hashimoto S, Kimura RS (1988) Computer-aided three-dimensional reconstruction and morphometry of the outer hair cells of the guinea pig cochlea. Acta Otolaryngol (Stockh) 105: 64–74

    Google Scholar 

  • Heizmann CW, Hunziker W (1991) Intracellular calcium-binding proteins: more sites than insights. Trends Biochem Sci 16: 98–103

    Google Scholar 

  • Howard J, Roberts WM, Hudspeth AJ (1988) Mechanoelectrical transduction by hair cells. Annu Rev Biophys Chem 17: 99–124

    Google Scholar 

  • Ichimiya I, Adams JC, Kimura RS (1994a) Immunolocalization of Na%#x002B;, K%#x002B;-ATPase, Ca%#x002B;+-ATPase, calcium-binding proteins, and carbonic anhydrase in the guinea pig inner ear. Acta Otolaryngol (Stockh) 114: 167–176

    Google Scholar 

  • Ichimiya I, Adams JC, Kimura RS (1994b) Changes in immunostaining of cochlea with experimentally induced endolymphatic hydrops. Ann Otol Rhinol Laryngol 103: 457–468

    Google Scholar 

  • Kikuchi T, Adams JC, Kimura RS (1995) Gap junctions in the rat cochlea: immunochemical and ultrastructural analysis. Anat Embryol 191: 101–118

    Google Scholar 

  • Kimitsuki T, Ohmori H (1992) The effect of caged calcium release on the adaptation of the transduction current in chick hair cells. J Physiol 458: 27–40

    Google Scholar 

  • Liberman MC (1978) Auditory-nerve response from cats raised in a low-noise chamber. J Acoust Soc Am 63: 442–455

    CAS  PubMed  Google Scholar 

  • Liberman MC, Dodds LW, Pierce S (1990) Afferent and efferent innervation of the cat cochlea; quantitative analysis with light and electron microscopy. J Comp Neurol 301: 443–460

    Google Scholar 

  • Lim DJ (1986) Functional structure of the organ of Corti: a review. Hear Res 22: 117–146

    Article  CAS  PubMed  Google Scholar 

  • Maurer J, Heinrich UR, Mann W (1994) Differences of inner and outer hair cells in the organ of Corti of the guinea pig in respect to the cellular content of precipitable calcium. Hear Res 72: 135–142

    Google Scholar 

  • Ohmori H (1985) Mechano-electrical transduction currents in isolated vestibular hair cells of the chick. J Physiol 359: 180–217

    Google Scholar 

  • Pack AK, Slepecky NB (1995) Comparison of the organization of cytoskeletal and calcium binding proteins in the mammalian organ of Corti: presence of cell specific proteins and longitudinal gradients. Unpublished abstracts of the 1995 Midwinter Meeting of the Association for Research in Otolaryngology, p 161

  • Pae SS, Saunders JC (1994) Intra- and extracellular calcium modulates stereocilia stiffness on chick hair cells. Proc Natl Acad Sci USA 91: 1153–1157

    Google Scholar 

  • Rabie A, Thomasset M, Legrand CC (1983) Immunocytochemical detection of calcium-binding protein in the cochlear and vestibular hair cells of the rat. Cell Tissue Res 232: 691–696

    Google Scholar 

  • Raymond J, Dechesne CJ, Desmadryl G, Dememes D (1993) Different calcium-binding proteins identify subpopulations of vestibular ganglion neurons in the rat. Acta Otolaryngol Suppl (Stockh) 503: 114–118

    Google Scholar 

  • Roberts WM, Howard J, Hudspeth A (1988) Hair cells: transduction, tuning and transmission in the inner ear. Ann Rev Cell Biol 4: 63–92

    Google Scholar 

  • Rogers JH (1989) Two calcium-binding proteins mark many chick sensory neurons. Neuroscience 31: 697–709

    Google Scholar 

  • Sacks DB, Porter SE, Ladenson JH, McDonald JM (1991) Monoclonal antibody to calmodulin: development, characterization, and comparison with polyclonal anti-calmodulin antibodies. Anal Biochem 194: 369–377

    Google Scholar 

  • Saito K (1983) Fine structure of the sensory epithelium of guinea-pig organ of Corti: subsurface cisternae and lamellar bodies in the outer hair cells. Cell Tissue Res 229: 467–481

    Google Scholar 

  • Sangameswaran L, Hempstead J, Morgan JI (1989) Molecular cloning of a neuron-specific transcript and its regulation during normal and aberrant cerebellar development. Proc Natl Acad Sci USA 86: 5651–5655

    Google Scholar 

  • Sans A, Etchecopar B, Brehier A, Thomasset M (1986) Immunocytochemical detection of vitamin D-dependent calcium-binding protein (CaBP-28K) in vestibular sensory hair cells and vestibular ganglion neurones of the cat. Brain Res 364: 190–194

    Google Scholar 

  • Sans A, Brehier A, Moniot B, Thomasset M (1987) Immuno-electromicroscopic localization of ‘vitamin D-dependent’ calcium-binding protein (CaBP-28k) in the vestibular hair cells of the cat. Brain Res 435: 293–304

    Google Scholar 

  • Schulte BA (1993) Immunohistochemical localization of intracellular Ca-ATPase in outer hair cells, neurons and fibrocytes in the adult and developing inner ear. Hear Res 65: 262–273

    Article  CAS  PubMed  Google Scholar 

  • Shepherd GMG, Barres BA, Corey DP (1989) “Bundle blot” purification and initial protein characterization of hair cell stereocilia. Proc Natl Acad Sci USA 86: 4973–4977

    PubMed  Google Scholar 

  • Slepecky NB, Ulfendahl M (1993) Evidence for calcium-binding proteins and calcium-dependent regulatory proteins in sensory cells of the organ of Corti. Hear Res 70: 73–84

    Article  CAS  PubMed  Google Scholar 

  • Slepecky NB, Ulfendahl M, Flock A (1988) Effect of caffeine and tetracaine on outer hair cell shortening suggest intracellular calcium involvement. Hear Res 32: 11–22

    Google Scholar 

  • Spicer SS, Schulte BA (1991) Differentiation of inner ear fibrocytes according to their ion transport related activity. Hear Res 56: 53–64

    Google Scholar 

  • Sridhar TS, Liberman MC, Brown MC, Sewell WF (1995a) A novel cholinergic “slow effect” of efferent stimulation of cochlear potentials in the guinea pig. J Neurosci 15: 3667–3678

    Google Scholar 

  • Sridhar TS, Brown MC, Sewell WF (1995b) Molecular mechanisms involved in olivocochlear efferent slow effects. Unpublished abstracts of the 1995 Midwinter Meeting of the Association for Research in Otolaryngology, p 172

  • Wasserman RH, Taylor AN (1966) Vitamin D3-induced calcium-binding protein.in chick intestinal mucosa. Science 152: 791–793

    CAS  Google Scholar 

  • Winsky L, Nakata H, Martin BM, Jacobowitz DM (1989) Isolation, partial amino acid sequence, and immunohistochemical localization of a brain calcium-binding protein. Proc Natl Acad Sci USA 86: 10139–10143

    Google Scholar 

  • Zenner HP (1988) Motility of outer hair cells as an active, actin-mediated process. Acta Otolaryngol (Stockh) 105: 39–44

    Google Scholar 

  • Ziai R, Pan YCE, Hulmes JD, Sangameswaran L, Morgan JI (1986) Isolation, sequence, and developmental profile of a brain-specific polypeptide, PEP-19. Proc Natl Acad Sci USA 83: 8420–8423

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Joe C. Adams.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Imamura, SI., Adams, J.C. Immunolocalization of peptide 19 and other calcium-binding proteins in the guinea pig cochlea. Anat Embryol 194, 407–418 (1996). https://doi.org/10.1007/BF00198543

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00198543

Key words

Navigation