bioRxiv preprint doi: https://doi.org/10.1101/396630; this version posted September 25, 2018. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY 4.0 International license. 1 Article 2 Zebrafish Otolith Biomineralization Requires 3 Polyketide Synthase 4 Kevin D. Thiessen 1, Steven J. Grzegorski 2, Lisa Higuchi 1, Jordan A. Shavit 2, and Kenneth L. 5 Kramer 1,* 6 1 Department of Biomedical Sciences, Creighton University School of Medicine, Omaha, NE, United States 7 2 Department of Pediatrics, University of Michigan, Ann Arbor, MI, United States 8 * Correspondence:
[email protected]; Tel.: 402-280-2763 9 Abstract: Deflecting biomineralized crystals attached to vestibular hair cells are necessary for 10 maintaining balance. Zebrafish (Danio rerio) are useful organisms to study these biomineralized 11 crystals called otoliths, as many required genes are homologous to human otoconial development. 12 We sought to identify and characterize the causative gene in a trio of mutants, no content (nco) and 13 corkscrew (csr), and vanished (vns), which fail to develop otoliths during early ear development. We 14 show that nco, csr, and vns have potentially deleterious mutations in polyketide synthase (pks1), a 15 multi-modular protein that has been previously implicated in biomineralization events in chordates 16 and echinoderms. We found that Otoconin-90 (Oc90) expression within the otocyst is normal in nco 17 and csr; therefore, it is not sufficient for otolith biomineralization in zebrafish. Similarly, normal 18 localization of Otogelin, a protein required for otolith tethering in the otolithic membrane, is not 19 sufficient for Oc90 attachment.