Abstract
Canonical transient receptor potential (TRPC) subunits assemble as tetramers to form ion channels with high calcium (Ca2+) permeability. Here, we investigated the possibility that TRPC3 ion channels are broadly expressed in the adult guinea pig and mouse cochleae. Using immunofluorescence, pronounced labeling occurred in the spiral ganglion (SG) neurons, inner hair cells (IHC), outer hair cells (OHC) and epithelial cells lining scala media. TRPC3 expression was homogeneous in the SG throughout the cochlea. In contrast, there was marked spatial variation in the immunolabeling in the cochlear hair cells with respect to location. This likely relates to the tonotopy of these cells. TRPC3 immunolabeling was more pronounced in the IHC than OHC. Both basal region IHC and OHC had higher TRPC3 expression levels than the corresponding cells from the apical region of the cochlea. These data suggest that TRPC3 ion channels contribute to Ca2+ homeostasis associated with the hair cells, with higher ion fluxes in more basal regions of the cochlea, and may also be a significant pathway for Ca2+ entry associated with auditory neurotransmission via the SG neurons. TRPC3 expression was also identified within the spiral limbus region, inner and outer sulcus, but without evidence for spatial variation in expression level. Expression in these gap junction-coupled epithelial cells lining scala media is indicative of a contribution of TRPC3 channels to cochlear electrochemical homeostasis.
Similar content being viewed by others
References
Becker EB, Oliver PL, Glitsch MD, Banks GT, Achilli F, Hardy A, Nolan PM, Fisher EM, Davies KE (2009) A point mutation in TRPC3 causes abnormal Purkinje cell development and cerebellar ataxia in moonwalker mice. PNAS (USA) 106:6706–6711
Chiba T, Marcus DC (2000) Nonselective cation and BK channels in apical membrane of outer sulcus epithelial cells. J Membr Biol 174:167–179
Corey DP (2006) What is the hair cell transduction channel? J Physiol 576:23–28
Corey DP, Garcia-Anoveros J, Holt JR, Kwan KY, Lin S-Y, Vollrath MA, Amalfitano A, Cheung ELM, Derfler BH, Duggan A, Geleoc GSG, Gray PA, Hoffman MP, Rehm HL, Tamasauskas D, Zhang D-S (2004) TRPA1 is a candidate for the mechanosensitive transduction channel of vertebrate hair cells. Nature 432:723–730
Cuajungco MP, Grimm C, Heller S (2007) TRP channels as candidates for hearing and balance abnormalities in vertebrates. Biochim Biophys Acta 1772:1022–1027
Damann N, Voets T, Nilius B (2008) TRPs in our senses. Curr Biol 18:R880–R889
Dulon D, Jagger DJ, Lin X, Davis RL (2006) Neuromodulation in the spiral ganglion: Shaping signals from the organ of Corti to the CNS. J Membr Biol 209:167–175
Friedman RA, Van Laer L, Huentelman MJ, Sheth SS, Van Eyken E, Corneveaux JJ, Tembe WD, Halperin RF, Thorburn AQ, Thys S, Bonneux S, Fransen E, Huyghe J, Pyykko I, Cremers CW, Kremer H, Dhooge I, Stephens D, Orzan E, Pfister M, Bille M, Parving A, Sorri M, Van de Heyning PH, Makmura L, Ohmen JD, Linthicum FH Jr, Fayad JN, Pearson JV, Craig DW, Stephan DA, Van Camp G (2009) GRM7 variants confer susceptibility to age-related hearing impairment. Hum Mol Genet 18:785–796
Hartmann J, Dragicevic E, Adelsberger H, Henning HA, Sumser M, Abramowitz J, Blum R, Dietrich A, Freichel M, Flockerzi V, Birnbaumer L, Konnerth A (2008) TRPC3 channels are required for synaptic transmission and motor coordination. Neuron 59:392–398
Housley GD, Marcotti W, Navaratnam D, Yamoah EN (2006) Hair cells-beyond the transducer. J Membr Biol 209:89–118
Huang WC, Young JS, Glitsch MD (2007) Changes in TRPC channel expression during postnatal development of cerebellar neurons. Cell Calcium 42:1–10
Ito K, Dulon D (2002) Nonselective cation conductance activated by muscarinic and purinergic receptors in rat spiral ganglion neurons. Am J Physiol Cell Physiol 282:C1121–C1135
Ito K, Rome C, Bouleau Y, Dulon D (2002) Substance P mobilizes intracellular calcium and activates a nonselective cation conductance in rat spiral ganglion neurons. Eur J Neurosci 16:2095–2102
Jagger DJ, Forge A (2006) Compartmentalized and signal-selective gap junctional coupling in the hearing cochlea. J Neurosci 26:1260–1268
Jia Y, Zhou J, Tai Y, Wang Y (2007) TRPC channels promote cerebellar granule neuron survival. Nat Neurosci 10:59–567
Kitahara T, Li H-S, Balaban CD (2005) Changes in transient receptor potential cation channel superfamily V (TRPV) mRNA expression in the mouse inner ear ganglia after kanamycin challenge. Hear Res 201:132–144
Li HS, Xu XZ, Montell C (1999) Activation of a TRPC3-dependent cation current through the neurotrophin BDNF. Neuron 24:261–273
Li Y, Jia YC, Cui K, Li N, Zheng ZY, Wang YZ, Yuan XB (2005) Essential role of TRPC channels in the guidance of nerve growth cones by brain-derived neurotrophic factor. Nature 434:894–898
Liang F, Niedzielski A, Schulte BA, Spicer SS, Hazen-Martin DJ, Shen Z (2003) A voltage- and Ca2+-dependent big conductance K channel in cochlear spiral ligament fibrocytes. Pflugers Arch Eur J Physiol 445:683–692
Liang F, Hu W, Schulte BA, Mao C, Qu C, Hazen-Martin DJ, Shen Z (2004) Identification and characterization of an L-type Cav1.2 channel in spiral ligament fibrocytes of gerbil inner ear. Brain Res Mol Brain Res 125:40–46
Mammano F, Frolenkov GI, Lagostena L, Belyantseva IA, Kurc M, Dodane V, Colavita A, Kachar B (1999) ATP-induced Ca2+ release in cochlear outer hair cells: localization of an inositol triphosphate-gated Ca2+ store to the base of the sensory hair bundle. J Neurosci 19:6918–6929
McGuinness SL, Shepherd RK (2005) Exogenous BDNF rescues rat spiral ganglion neurons in vivo. Otol Neurotol 26:1064–1072
Mou K, Adamson CL, Davis RL (1998) Time-dependence and cell-type specificity of synergistic neurotrophin actions on spiral ganglion neurons. J Comp Neurol 402:129–139
O’Neil RG, Heller S (2005) The mechanosensitive nature of TRPV channels. Pflugers Arch Eur J Physiol 451:193–203
Peng BG, Li QX, Ren TY, Ahmad S, Chen SP, Chen P, Lin X (2004) Group I metabotropic glutamate receptors in spiral ganglion neurons contribute to excitatory neurotransmissions in the cochlea. Neuroscience 123:221–230
Ramsey IS, Delling M, Clapham DE (2006) An introduction to TRP channels. Ann Rev Physiol 68:619–647
Raybould NP, Housley GD (1997) Variation in expression of the outer hair cell P2X receptor conductance along the guinea-pig cochlea. J Physiol 498:717–727
Raybould NP, Jagger DJ, Housley GD (2001) Positional analysis of guinea-pig inner hair cell membrane conductances: implications for regulation of the membrane filter. JARO 2:362–376
Raybould NP, Jagger DJ, Kanjhan R, Greenwood D, Laslo P, Hoya N, Soeller C, Cannell MB, Housley GD (2007) TRPC-like conductance mediates restoration of intracellular Ca2+ in cochlear outer hair cells in the guinea-pig and rat. J Physiol 579:101–113
Roehm PC, Xu N, Woodson EA, Green SH, Hansen MR (2008) Membrane depolarization inhibits spiral ganglion neurite growth via activation of multiple types of voltage sensitive calcium channels and calpain. Mol Cell Neurosci 37:376–387
Shen Z, Liang F, Hazen-Martin DJ, Schulte BA (2004) BK channels mediate the voltage-dependent outward current in type I spiral ligament fibrocytes. Hear Res 187:35–43
Shen J, Harada N, Kubo N, Liu B, Mizuno A, Suzuki M, Yamashita T (2006) Functional expression of transient receptor potential vanilloid 4 in the mouse cochlea. Neuroreport 17:135–139
Skinner LJ, Enee V, Beurg M, Jung HH, Ryan AF, Hafidi A, Aran JM, Dulon D (2003) Contribution of BK Ca2+-activated K+ channels to auditory neurotransmission in the Guinea-pig cochlea. J Neurophysiol 90:320–332
Spicer SS, Schulte BA (1998) Evidence for a medial K+ recycling pathway from inner hair cells. Hear Res 118:1–12
Sun J, Ahmad S, Chen S, Tang W, Zhang Y, Chen P, Lin X (2005) Cochlear gap junctions coassembled from Cx26 and 30 show faster intercellular Ca2+ signaling than homomeric counterparts. Am J Physiol 288:C613–C623
Tabuchi K, Suzuki M, Mizuno A, Hara A (2005) Hearing impairment in TRPV4 knockout mice. Neurosci Lett 382:304–308
Tang J, Lin Y, Zhang Z, Tikunova S, Birnbaumer L, Zhu MX (2001) Identification of common binding sites for calmodulin and inositol 1, 4, 5-trisphosphate receptors on the carboxyl termini of trp channels. J Biol Chem 276:21303–21310
van Aken AFJ, Atiba-Davies M, Marcotti W, Goodyear RJ, Bryant JE, Richardson GP, Noben-Trauth K, Kros CJ (2008) TRPML3 mutations cause impaired mechano-electrical transduction and depolarization by an inward-rectifier cation current in auditory hair cells of varitint-waddler mice. J Physiol 586:5403–5418
Ylikoski J, Pirvola U, Moshnyakov M, Palgi J, Arumae U, Saarma M (1993) Expression patterns of neurotrophin and their receptor mRNAs in the rat inner ear. Hear Res 65:69–78
Zhang Z, Tang J, Tikunova S, Johnson JD, Chen Z, Qin N, Dietrich A, Stefani E, Birnbaumer L, Zhu MX (2001) Activation of Trp3 by inositol 1, 4, 5-trisphosphate receptors through displacement of inhibitory calmodulin from a common binding domain. PNAS (USA) 98:3168–3173
Acknowledgments
We acknowledge support from UNSW Faculty of Medicine Translational Neuroscience Facility funding and in part, the support of the Intramural Research Program of the NIH (Project Z01-ES101684 to LB).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Tadros, S.F., Kim, Y., Phan, P.A.B. et al. TRPC3 ion channel subunit immunolocalization in the cochlea. Histochem Cell Biol 133, 137–147 (2010). https://doi.org/10.1007/s00418-009-0653-6
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-009-0653-6