Identification of the hair cell soma-1 antigen, HCS-1, as otoferlin.
Hair cells, the mechanosensitive receptor cells of the inner ear, are critical for our senses of hearing and balance. The small number of these receptor cells in the inner ear has impeded the identification and characterization of proteins important for hair cell function. The binding specificity of monoclonal antibodies provides a means for identifying hair cell-specific proteins and isolating them for further study. We have generated a monoclonal antibody, termed hair cell soma-1 (HCS-1), which specifically immunolabels hair cells in at least five vertebrate classes, including sharks and rays, bony fish, amphibians, birds, and mammals. We used HCS-1 to immunoprecipitate the cognate antigen and identified it as otoferlin, a member of the ferlin protein family. Mutations in otoferlin underlie DFNB9, a recessive, nonsyndromic form of prelingual deafness characterized as an auditory neuropathy. Using immunocytochemistry, we find that otoferlin is associated with the entire basolateral membrane of the hair cells and with vesicular structures distributed throughout most of the hair cell cytoplasm. Biochemical assays indicate that otoferlin is tightly associated with membranes, as it is not solubilized by alterations in calcium or salt concentrations. HCS-1 immunolabeling does not co-localize with ribeye, a constituent of synaptic ribbons, suggesting that otoferlin may, in addition to its proposed function in synaptic vesicle release, play additional roles in hair cells.
Goodyear
Richard J
RJ
School of Life Sciences, University of Sussex, Falmer, Brighton, BN1 9QG, UK. r.j.goodyear@sussex.ac.uk
Legan
P Kevin
PK
Christiansen
Jeffrey R
JR
Xia
Bei
B
Korchagina
Julia
J
Gale
Jonathan E
JE
Warchol
Mark E
ME
Corwin
Jeffrey T
JT
Richardson
Guy P
GP
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000378:metabolism; D002645:Chickens; D003638:Deafness / Q000235:genetics / Q000378:metabolism; D004284:Dogfish; D005849:Gerbillinae; D006168:Guinea Pigs; D006199:Hair Cells, Auditory, Inner / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D023421:Models, Animal; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D011892:Rana catesbeiana; D013572:Synaptic Vesicles / Q000378:metabolism; D015027:Zebrafish
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins
J Med Genet. 2002 Jul;39(7):502-6
12114484
Hum Mol Genet. 2009 Aug 1;18(15):2779-90
19417007
Genes Dev. 2007 Sep 1;21(17):2118-30
17785522
J Neurosci. 1999 Jan 1;19(1):119-32
9870944
Cell. 2006 Oct 20;127(2):277-89
17055430
Dev Biol. 2006 Jun 1;294(1):148-60
16566913
Proc Natl Acad Sci U S A. 1995 Oct 24;92(22):10297-301
7479771
Hum Mutat. 2007 Jun;28(6):571-7
17301963
Hum Mutat. 2003 Dec;22(6):451-6
14635104
Int J Dev Neurosci. 1997 Jul;15(4-5):417-32
9263023
Proc Natl Acad Sci U S A. 2005 Feb 1;102(5):1496-501
15677337
Nat Rev Mol Cell Biol. 2008 Feb;9(2):99-111
18216767
Neurobiol Dis. 2002 Jul;10(2):157-64
12127154
J Comp Neurol. 2007 Feb 1;500(4):646-57
17154269
J Neurosci. 1997 Aug 15;17(16):6289-301
9236239
J Med Genet. 2006 Jul;43(7):576-81
16371502
J Neurosci. 2006 Oct 11;26(41):10438-51
17035528
J Comp Neurol. 2000 Jan 31;417(1):1-16
10660884
J Neurosci. 2006 Jun 28;26(26):7022-34
16807332
Brain Res Brain Res Protoc. 2002 Feb;9(1):65-76
11852272
J Biol Chem. 2009 Jan 16;284(3):1364-72
19004828
J Neurosci. 2006 Jun 14;26(24):6543-53
16775142
Development. 2005 Feb;132(3):541-51
15634704
Hear Res. 2007 Dec;234(1-2):21-8
17967520
J Neurosci. 2003 Jun 15;23(12):4878-87
12832510
Biochim Biophys Acta. 2006 Aug;1761(8):838-49
16945584
J Neurosci. 2008 Feb 27;28(9):2261-73
18305259
Am J Med Genet A. 2005 Sep 15;138(1):6-10
16097006
J Neurosci. 2007 Feb 28;27(9):2163-75
17329413
J Neurobiol. 2002 Feb 5;50(2):81-92
11793356
J Cell Biol. 1990 Apr;110(4):1055-66
2182645
J Med Genet. 2003 Jan;40(1):45-50
12525542
J Comp Neurol. 1999 Apr 5;406(2):183-98
10096605
Cell. 1988 Feb 12;52(3):329-41
2964276
J Neurosci. 2003 Oct 8;23(27):9208-19
14534255
J Neurosci. 2007 Mar 21;27(12):3174-86
17376979
Proc Natl Acad Sci U S A. 2007 Oct 16;104(42):16675-80
17895386
Trends Cell Biol. 2004 Apr;14(4):206-13
15066638
J Neurosci. 2008 Feb 20;28(8):1798-803
18287496
Am J Hum Genet. 2000 Sep;67(3):591-600
10903124
Clin Genet. 2009 Mar;75(3):237-43
19250381
Neuron. 2005 Nov 3;48(3):403-15
16269359
J Comp Neurol. 2005 Feb 28;483(1):91-113
15672394
Hear Res. 1989 Nov;42(2-3):175-94
2606802
J Neurosci. 2010 Sep 15;30(37):12545-56
20844149
Nat Genet. 1999 Apr;21(4):363-9
10192385
J Neurosci. 2001 Jul 15;21(14):5066-78
11438582
J Assoc Res Otolaryngol. 2000 Sep;1(2):172-82
11545144
Eur J Neurosci. 2006 Dec;24(12):3372-80
17229086
J Neurocytol. 1996 Apr;25(4):257-66
8793731
Neuron. 2000 Dec;28(3):857-72
11163272
Proc Natl Acad Sci U S A. 2006 Dec 5;103(49):18615-20
17124170
Nature. 1989 Oct 5;341(6241):397-404
2677742
J Neurosci. 1999 May 15;19(10):3761-72
10234008
J Comp Neurol. 1992 Nov 8;325(2):243-56
1281174
J Comp Neurol. 1991 Dec 22;314(4):777-88
1816275
J Comp Neurol. 2005 Mar 28;484(1):105-20
15717301