Novel roles for the radial spoke head protein 9 in neural and neurosensory cilia.
Cilia are cell surface organelles with key roles in a range of cellular processes, including generation of fluid flow by motile cilia. The axonemes of motile cilia and immotile kinocilia contain 9 peripheral microtubule doublets, a central microtubule pair, and 9 connecting radial spokes. Aberrant radial spoke components RSPH1, 3, 4a and 9 have been linked with primary ciliary dyskinesia (PCD), a disorder characterized by ciliary dysmotility; yet, radial spoke functions remain unclear. Here we show that zebrafish Rsph9 is expressed in cells bearing motile cilia and kinocilia, and localizes to both 9 + 2 and 9 + 0 ciliary axonemes. Using CRISPR mutagenesis, we show that rsph9 is required for motility of presumptive 9 + 2 olfactory cilia and, unexpectedly, 9 + 0 neural cilia. rsph9 is also required for the structural integrity of 9 + 2 and 9 + 0 ciliary axonemes. rsph9 mutant larvae exhibit reduced initiation of the acoustic startle response consistent with hearing impairment, suggesting a novel role for Rsph9 in the kinocilia of the inner ear and/or lateral line neuromasts. These data identify novel roles for Rsph9 in 9 + 0 motile cilia and in sensory kinocilia, and establish a useful zebrafish PCD model.
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.
Irina
I
Sedykh
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.|Cellular and Molecular Biology Training Program, University of Wisconsin, Madison, WI, 53706, USA.
Jessica J
JJ
TeSlaa
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.
Rose L
RL
Tatarsky
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.
Abigail N
AN
Keller
Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI 53706, USA.|McPherson Eye Research Institute, University of Wisconsin, Madison, WI, 53706, USA.
Kimberly A
KA
Toops
Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Madison, WI 53706, USA.|McPherson Eye Research Institute, University of Wisconsin, Madison, WI, 53706, USA.
Aparna
A
Lakkaraju
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.
Molly K
MK
Nyholm
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.
Marc A
MA
Wolman
Department of Zoology, University of Wisconsin, Madison, WI, 53706, USA.|Department of Neuroscience, University of Wisconsin, Madison, WI, 53706, USA.|McPherson Eye Research Institute, University of Wisconsin, Madison, WI, 53706, USA.
Yevgenya
Y
Grinblat
J Exp Biol. 2005 Apr;208(Pt 7):1363-72
15781896
Nat Cell Biol. 2010 Apr;12(4):407-12
20305649
Neuroscience. 1982;7(12):3091-103
6984492
J Cell Biol. 1970 Oct;47(1):159-82
4935335
Am J Hum Genet. 2013 Oct 3;93(4):672-86
24094744
Curr Biol. 2014 Oct 6;24(19):R973-82
25291643
Hum Mol Genet. 2014 Jul 1;23(13):3362-74
24518672
Hum Mutat. 2013 Mar;34(3):462-72
23255504
J Comp Neurol. 1993 Jul 8;333(2):289-300
8345108
Dev Cell. 2015 Mar 23;32(6):756-64
25752963
Cells. 2015 Sep 11;4(3):500-19
26378583
BMC Dev Biol. 2006 Jan 13;6:1
16412219
Nat Commun. 2014 Dec 04;5:5727
25473808
Dev Dyn. 2007 Jul;236(7):1963-9
17503454
Am J Respir Cell Mol Biol. 2015 Oct;53(4):563-73
25789548
Methods Cell Biol. 2010;97:415-35
20719283
Cilia. 2015 Jan 22;4(1):2
25610612
Dev Dyn. 2003 Nov;228(3):464-74
14579384
J Cell Biol. 1985 Jun;100(6):2008-18
2860115
Dev Dyn. 1995 Jul;203(3):253-310
8589427
J Cell Biol. 1981 Dec;91(3 Pt 2):107s-124s
6459326
J Otolaryngol. 2002 Feb;31(1):13-7
11881766
Development. 2014 Apr;141(7):1427-41
24644260
Am J Respir Crit Care Med. 2014 Mar 15;189(6):707-17
24568568
PLoS One. 2013;8(3):e59436
23527195
Nat Genet. 2000 Oct;26(2):216-20
11017081
Development. 2012 May;139(10):1777-87
22461562
Proc Natl Acad Sci U S A. 2010 Oct 26;107(43):18499-504
20937855
J Neurosci. 2007 May 2;27(18):4984-94
17475807
Nat Rev Genet. 2010 May;11(5):331-44
20395968
Am J Hum Genet. 2013 Sep 5;93(3):561-70
23993197
Development. 2005 Apr;132(8):1907-21
15790966
Annu Rev Physiol. 2007;69:401-22
16945069
Chem Senses. 1998 Feb;23(1):39-48
9530968
PLoS One. 2012;7(3):e33667
22448264
Dev Biol. 2008 Feb 15;314(2):261-75
18178183
Hum Mol Genet. 2015 May 1;24(9):2482-91
25601850
J Med Genet. 2014 Jan;51(1):61-7
24203976
Dev Cell. 2015 Oct 26;35(2):236-46
26506310
Nature. 2009 Jan 8;457(7226):205-9
19043402
J Biophys Biochem Cytol. 1959 Mar 25;5(2):269-78
13654448
Hum Mol Genet. 2002 Mar 15;11(6):715-21
11912187
Nat Rev Drug Discov. 2014 Oct;13(10):759-80
25233993
Hum Mol Genet. 2009 Jan 15;18(2):289-303
18971206
Nat Biotechnol. 2013 Mar;31(3):227-9
23360964
Nature. 2009 May 7;459(7243):98-102
19305393
Nat Genet. 2008 Dec;40(12):1445-53
19011630
Am J Hum Genet. 2013 Aug 8;93(2):346-56
23891471
PLoS One. 2008 Sep 01;3(9):e3090
18769618
Am J Respir Crit Care Med. 2013 Oct 15;188(8):913-22
23796196
Annu Rev Physiol. 2007;69:377-400
17009929
Development. 2009 Nov;136(22):3791-800
19855021
Proc Natl Acad Sci U S A. 2011 Sep 13;108(37):15468-73
21876167
Development. 2005 Mar;132(6):1247-60
15716348
Biol Open. 2012 Aug 15;1(8):815-25
23213475
J Cell Biol. 1974 Oct;63(1):35-63
4424314
Bioarchitecture. 2014 Jan-Feb;4(1):6-15
24481178
Methods Cell Biol. 2009;93:197-217
20409819
Am J Hum Genet. 2013 Oct 3;93(4):711-20
24055112
Am J Hum Genet. 2013 Aug 8;93(2):357-67
23849778
Development. 2014 Sep;141(17):3410-9
25139857
Genet Med. 2009 Jul;11(7):473-87
19606528
PLoS One. 2011;6(5):e19713
21603650
Development. 2004 Aug;131(16):4085-93
15269167
J Cell Biol. 1965 Apr;25:1-8
14283628
PLoS One. 2013 Aug 26;8(8):e72299
23991085
Dev Dyn. 2004 Jul;230(3):403-9
15188426
J Cell Biol. 2014 Mar 3;204(5):807-19
24590175
Hum Mol Genet. 2004 Sep 15;13(18):2133-41
15269178
Am J Hum Genet. 2009 Feb;84(2):197-209
19200523
Cell. 2006 Apr 7;125(1):33-45
16615888
Methods Enzymol. 2013;525:219-44
23522472
Am J Hum Genet. 2015 Jul 2;97(1):153-62
26073779
Biotechniques. 2007 Nov;43(5):610, 612, 614
18072590