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Thiessen KD
,
Grzegorski SJ
,
Chin Y
,
Higuchi LN
,
Wilkinson CJ
,
Shavit JA
,
Kramer KL
.
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Deflecting biomineralized crystals attached to vestibular hair cells are necessary for maintaining balance. Zebrafish (Danio rerio) are useful organisms to study these biomineralized crystals called otoliths, as many required genes are homologous to human otoconial development. We sought to identify and characterize the causative gene in a trio of homozygous recessive mutants, no content (nco) and corkscrew (csr), and vanished (vns), which fail to develop otoliths during early ear development. We show that nco, csr, and vns have potentially deleterious mutations in polyketide synthase (pks1), a multi-modular protein that has been previously implicated in biomineralization events in chordates and echinoderms. We found that Otoconin-90 (Oc90) expression within the otocyst is diffuse in nco and csr; therefore, it is not sufficient for otolith biomineralization in zebrafish. Similarly, normal localization of Otogelin, a protein required for otolith tethering in the otolithic membrane, is not sufficient for Oc90 attachment. Furthermore, eNOS signaling and Endothelin-1 signaling were the most up- and down-regulated pathways during otolith agenesis in nco, respectively. Our results demonstrate distinct processes for otolith nucleation and biomineralization in vertebrates and will be a starting point for models that are independent of Oc90-mediated seeding. This study will serve as a basis for investigating the role of eNOS signaling and Endothelin-1 signaling during otolith formation.
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30974150
???displayArticle.pmcLink???PMC6531356 ???displayArticle.link???Mech Dev ???displayArticle.grants???[+]
Figure 2:. Complementary approaches for causative gene discovery. MMAPPR analysis of RNA sequencing data for nco (A) and whole genome homology mapping for csr (B) identified regions of high homology on the 24th chormosome near the pks1 locus (~33 Gb). (C) Deleterious mutations were identified in pks1 for nco and csr within the acyl transferase (AT) domain and vns within the polyketide synthase (PKS) domain. Sanger sequencing confirmed SNPs in csr, nco, and vns mutants. Other domains include Ketoacyl Synthetase (KS), Medium Chain Reductase (MDR), NAD(P)-dependent dehydrogenase (NDD), and Phosphopanthetheine-Binding (PP).
Figure 3:. WT pks1 nucleic acid rescues otolith formation in csr, nco, and vns. (A) Normal frequencies of mutant phenotypes in each uninjected strain. All four pairings follow homozygous recessive mode of inheritance. (B) Results of injected embryos show that Japanese medaka pks1 mRNA (300 pg) rescues both csr and nco mutants and pks1 DNA (20 pg) rescues vns mutants. (*, p < 0.0001, paired t-test)(**, p < 0.0032, paired t-test)(***, p = 0.0001, paired t-test),. Site-directed mutagenesis was used to introduce a conserved mutation in csr (A911P) into the Japanese medaka construct (L905P) (C) Injection of pks1L905P (300 pg) fails to rescue csr or nco mutant phenotypes.
Figure 4:. Gene expression and pathway analysis of nco embryos. (A) Ingenuity Pathway Analysis shows the top up-regulated and down-regulated pathways, which are eNOS Signaling and Endothelin-1 Signaling, respectively. Positive z-score indicated increased mRNA levels. Negative z-score indicates decreased mRNA levels. No change in mRNA levels results in a z-score of zero. (B) Differential gene expression in the top up-regulated genes. (C) Differential gene expression in the top down-regulated genes. (**, expressed in adult zebrafish mechanosensory hair cells) [32].
Figure 5:. Aberrant expression of proteins invovled in otolith development in csr and nco. (A) Schematic of anterior macula (AM) tethered to otolith at 27 hpf. (B) In WT, Otoconin-90 (Oc90) is expressed within the mineralized otolith, which is situated atop the otolithic membrane (Otogelin, or Otog), at 27 hpf. Scale bar = 5 μm. (C-D) Oc90 has diffuse expression within the otocyst of csr and nco. In csr and nco, Otog is localized near the apical surface of hair cells. (E-F) Expression showing hair cells in WT and nco larvae at 5dpf. Scale bar = 25 μm. (G) Quantification of hair cell numbers in the posterior and anterior macula of WT and nco (n = 4).
Aceto,
Zebrafish Bone and General Physiology Are Differently Affected by Hormones or Changes in Gravity.
2015, Pubmed
Aceto,
Zebrafish Bone and General Physiology Are Differently Affected by Hormones or Changes in Gravity.
2015,
Pubmed
Barta,
RNA-seq transcriptomic analysis of adult zebrafish inner ear hair cells.
2018,
Pubmed
Borelli,
Biochemical relationships between endolymph and otolith matrix in the trout (Oncorhynchus mykiss) and turbot (Psetta maxima).
2001,
Pubmed
Chang,
An enhanced computational platform for investigating the roles of regulatory RNA and for identifying functional RNA motifs.
2013,
Pubmed
Chung,
Structural and molecular interrogation of intact biological systems.
2013,
Pubmed
Colantonio,
The dynein regulatory complex is required for ciliary motility and otolith biogenesis in the inner ear.
2009,
Pubmed
Deans,
Mammalian Otolin: a multimeric glycoprotein specific to the inner ear that interacts with otoconial matrix protein Otoconin-90 and Cerebellin-1.
2010,
Pubmed
Esaki,
Mechanism of development of ionocytes rich in vacuolar-type H(+)-ATPase in the skin of zebrafish larvae.
2009,
Pubmed
Geisler,
Large-scale mapping of mutations affecting zebrafish development.
2007,
Pubmed
Haddon,
Early ear development in the embryo of the zebrafish, Danio rerio.
1996,
Pubmed
Hill,
MMAPPR: mutation mapping analysis pipeline for pooled RNA-seq.
2013,
Pubmed
Hojo,
Unexpected link between polyketide synthase and calcium carbonate biomineralization.
2015,
Pubmed
,
Echinobase
Hughes,
Otopetrin 1 is required for otolith formation in the zebrafish Danio rerio.
2004,
Pubmed
Jiang,
Gene-expression analysis of hair cell regeneration in the zebrafish lateral line.
2014,
Pubmed
Johnson,
Big endothelin changes the cellular miRNA environment in TMOb osteoblasts and increases mineralization.
2014,
Pubmed
Kirchmaier,
The genomic and genetic toolbox of the teleost medaka (Oryzias latipes).
2015,
Pubmed
Kiss,
Inactivation of NADPH oxidase organizer 1 results in severe imbalance.
2006,
Pubmed
Krämer,
Causal analysis approaches in Ingenuity Pathway Analysis.
2014,
Pubmed
Kumai,
A role for transcription factor glial cell missing 2 in Ca2+ homeostasis in zebrafish, Danio rerio.
2015,
Pubmed
Liu,
[The study on plasma ET and NO of patients with sudden hearing loss].
2003,
Pubmed
Lundberg,
Mechanisms of otoconia and otolith development.
2015,
Pubmed
Miller,
sucker encodes a zebrafish Endothelin-1 required for ventral pharyngeal arch development.
2000,
Pubmed
Mitchison,
Mutations in axonemal dynein assembly factor DNAAF3 cause primary ciliary dyskinesia.
2012,
Pubmed
Moreland,
In vitro calcite crystal morphology is modulated by otoconial proteins otolin-1 and otoconin-90.
2014,
Pubmed
Obholzer,
Rapid positional cloning of zebrafish mutations by linkage and homozygosity mapping using whole-genome sequencing.
2012,
Pubmed
Peterson,
Small molecule developmental screens reveal the logic and timing of vertebrate development.
2000,
Pubmed
Petko,
Otoc1: a novel otoconin-90 ortholog required for otolith mineralization in zebrafish.
2008,
Pubmed
Pingault,
Review and update of mutations causing Waardenburg syndrome.
2010,
Pubmed
Reimer,
Rapid growth causes abnormal vaterite formation in farmed fish otoliths.
2017,
Pubmed
Riley,
A critical period of ear development controlled by distinct populations of ciliated cells in the zebrafish.
1997,
Pubmed
Riley,
Genes controlling the development of the zebrafish inner ear and hair cells.
2003,
Pubmed
Riley,
A mutation in zebrafish affecting a localized cellular function required for normal ear development.
1996,
Pubmed
Schibler,
A screen for genetic defects of the zebrafish ear.
2007,
Pubmed
Stawicki,
The zebrafish merovingian mutant reveals a role for pH regulation in hair cell toxicity and function.
2014,
Pubmed
Stooke-Vaughan,
The role of hair cells, cilia and ciliary motility in otolith formation in the zebrafish otic vesicle.
2012,
Pubmed
Stooke-Vaughan,
Otolith tethering in the zebrafish otic vesicle requires Otogelin and α-Tectorin.
2015,
Pubmed
Sumanas,
Zebrafish chaperone protein GP96 is required for otolith formation during ear development.
2003,
Pubmed
Surup,
The iromycins, a new family of pyridone metabolites from Streptomyces sp. I. Structure, NOS inhibitory activity, and biosynthesis.
2007,
Pubmed
Söllner,
Control of crystal size and lattice formation by starmaker in otolith biomineralization.
2003,
Pubmed
Söllner,
Mutated otopetrin 1 affects the genesis of otoliths and the localization of Starmaker in zebrafish.
2004,
Pubmed
Thomas,
The inner ear proteome of fish.
2019,
Pubmed
Ulitsky,
Extensive alternative polyadenylation during zebrafish development.
2012,
Pubmed
Wang,
Otoconin-90, the mammalian otoconial matrix protein, contains two domains of homology to secretory phospholipase A2.
1998,
Pubmed
White,
A high-resolution mRNA expression time course of embryonic development in zebrafish.
2017,
Pubmed
Whitfield,
Mutations affecting development of the zebrafish inner ear and lateral line.
1996,
Pubmed
Yang,
Matrix recruitment and calcium sequestration for spatial specific otoconia development.
2011,
Pubmed
Yu,
Cilia-driven fluid flow as an epigenetic cue for otolith biomineralization on sensory hair cells of the inner ear.
2011,
Pubmed
Zhao,
Otoconin-90 deletion leads to imbalance but normal hearing: a comparison with other otoconia mutants.
2008,
Pubmed
Zhao,
Gene targeting reveals the role of Oc90 as the essential organizer of the otoconial organic matrix.
2007,
Pubmed
