Regionalized Protein Localization Domains in the Zebrafish Hair Cell Kinocilium

Overview

Journal J Dev Biol

Specialty Biology

Date 2023 Jun 27

PMID 37367482

Authors

Timothy Erickson

William Paul Biggers 3rd

Kevin Williams

Shyanne E Butland

Alexandra Venuto

Affiliations

Soon will be listed here.

Abstract

Sensory hair cells are the receptors for auditory, vestibular, and lateral line sensory organs in vertebrates. These cells are distinguished by "hair"-like projections from their apical surface collectively known as the hair bundle. Along with the staircase arrangement of the actin-filled stereocilia, the hair bundle features a single, non-motile, true cilium called the kinocilium. The kinocilium plays an important role in bundle development and the mechanics of sensory detection. To understand more about kinocilial development and structure, we performed a transcriptomic analysis of zebrafish hair cells to identify cilia-associated genes that have yet to be characterized in hair cells. In this study, we focused on three such genes-, , and -because human or mouse orthologs are either associated with sensorineural hearing loss or are located near uncharacterized deafness loci. We made transgenic fish that express fluorescently tagged versions of their proteins, demonstrating their localization to the kinocilia of zebrafish hair cells. Furthermore, we found that Ankef1a, Odf3l2a, and Saxo2 exhibit distinct localization patterns along the length of the kinocilium and within the cell body. Lastly, we have reported a novel overexpression phenotype of Saxo2. Overall, these results suggest that the hair cell kinocilium in zebrafish is regionalized along its proximal-distal axis and set the groundwork to understand more about the roles of these kinocilial proteins in hair cells.

References

Erickson T, Nicolson T . Identification of sensory hair-cell transcripts by thiouracil-tagging in zebrafish. BMC Genomics. 2015; 16:842. PMC: 4619078. DOI: 10.1186/s12864-015-2072-5. View

Sheets L, He X, Olt J, Schreck M, Petralia R, Wang Y . Enlargement of Ribbons in Zebrafish Hair Cells Increases Calcium Currents But Disrupts Afferent Spontaneous Activity and Timing of Stimulus Onset. J Neurosci. 2017; 37(26):6299-6313. PMC: 5490065. DOI: 10.1523/JNEUROSCI.2878-16.2017. View

Kindt K, Finch G, Nicolson T . Kinocilia mediate mechanosensitivity in developing zebrafish hair cells. Dev Cell. 2012; 23(2):329-41. PMC: 3426295. DOI: 10.1016/j.devcel.2012.05.022. View

Bowl M, Brown S . Genetic landscape of auditory dysfunction. Hum Mol Genet. 2018; 27(R2):R130-R135. DOI: 10.1093/hmg/ddy158. View

Groza T, Lopez Gomez F, Haseli Mashhadi H, Munoz-Fuentes V, Gunes O, Wilson R . The International Mouse Phenotyping Consortium: comprehensive knockout phenotyping underpinning the study of human disease. Nucleic Acids Res. 2022; 51(D1):D1038-D1045. PMC: 9825559. DOI: 10.1093/nar/gkac972. View

Schwenk B, Lang C, Hogl S, Tahirovic S, Orozco D, Rentzsch K . The FTLD risk factor TMEM106B and MAP6 control dendritic trafficking of lysosomes. EMBO J. 2013; 33(5):450-67. PMC: 3989627. DOI: 10.1002/embj.201385857. View

Stauber M, Weidemann M, Dittrich-Breiholz O, Lobschat K, Alten L, Mai M . Identification of FOXJ1 effectors during ciliogenesis in the foetal respiratory epithelium and embryonic left-right organiser of the mouse. Dev Biol. 2016; 423(2):170-188. DOI: 10.1016/j.ydbio.2016.11.019. View

Sheets L, Hagen M, Nicolson T . Characterization of Ribeye subunits in zebrafish hair cells reveals that exogenous Ribeye B-domain and CtBP1 localize to the basal ends of synaptic ribbons. PLoS One. 2014; 9(9):e107256. PMC: 4160224. DOI: 10.1371/journal.pone.0107256. View

Jenkinson E, Rehman A, Walsh T, Clayton-Smith J, Lee K, Morell R . Perrault syndrome is caused by recessive mutations in CLPP, encoding a mitochondrial ATP-dependent chambered protease. Am J Hum Genet. 2013; 92(4):605-13. PMC: 3617381. DOI: 10.1016/j.ajhg.2013.02.013. View

10.

Charizopoulou N, Lelli A, Schraders M, Ray K, Hildebrand M, Ramesh A . Gipc3 mutations associated with audiogenic seizures and sensorineural hearing loss in mouse and human. Nat Commun. 2011; 2:201. PMC: 3105340. DOI: 10.1038/ncomms1200. View

11.

Ross A, May-Simera H, Eichers E, Kai M, Hill J, Jagger D . Disruption of Bardet-Biedl syndrome ciliary proteins perturbs planar cell polarity in vertebrates. Nat Genet. 2005; 37(10):1135-40. DOI: 10.1038/ng1644. View

12.

Steiner A, Kim T, Cabot V, Hudspeth A . Dynamic gene expression by putative hair-cell progenitors during regeneration in the zebrafish lateral line. Proc Natl Acad Sci U S A. 2014; 111(14):E1393-401. PMC: 3986164. DOI: 10.1073/pnas.1318692111. View

13.

Egydio de Carvalho C, Tanaka H, Iguchi N, Ventela S, Nojima H, Nishimune Y . Molecular cloning and characterization of a complementary DNA encoding sperm tail protein SHIPPO 1. Biol Reprod. 2002; 66(3):785-95. DOI: 10.1095/biolreprod66.3.785. View

14.

Konno A, Shiba K, Cai C, Inaba K . Branchial cilia and sperm flagella recruit distinct axonemal components. PLoS One. 2015; 10(5):e0126005. PMC: 4427456. DOI: 10.1371/journal.pone.0126005. View

15.

Shen J, Scheffer D, Kwan K, Corey D . SHIELD: an integrative gene expression database for inner ear research. Database (Oxford). 2015; 2015:bav071. PMC: 4513695. DOI: 10.1093/database/bav071. View

16.

Erickson T, Pacentine I, Venuto A, Clemens R, Nicolson T . The Ohnologs and 5b Are Required for Mechanotransduction in Distinct Populations of Sensory Hair Cells in Zebrafish. Front Mol Neurosci. 2020; 12:320. PMC: 6974483. DOI: 10.3389/fnmol.2019.00320. View

17.

Baker M, Hetherington L, Reeves G, Aitken R . The mouse sperm proteome characterized via IPG strip prefractionation and LC-MS/MS identification. Proteomics. 2008; 8(8):1720-30. DOI: 10.1002/pmic.200701020. View

18.

Flock A, DUVALL 3rd A . THE ULTRASTRUCTURE OF THE KINOCILIUM OF THE SENSORY CELLS IN THE INNER EAR AND LATERAL LINE ORGANS. J Cell Biol. 1965; 25:1-8. PMC: 2106605. DOI: 10.1083/jcb.25.1.1. View

19.

Sobkowicz H, Slapnick S, August B . The kinocilium of auditory hair cells and evidence for its morphogenetic role during the regeneration of stereocilia and cuticular plates. J Neurocytol. 1995; 24(9):633-53. DOI: 10.1007/BF01179815. View

20.

Thomas P, Ebert D, Muruganujan A, Mushayahama T, Albou L, Mi H . PANTHER: Making genome-scale phylogenetics accessible to all. Protein Sci. 2021; 31(1):8-22. PMC: 8740835. DOI: 10.1002/pro.4218. View