Separate Na,K-ATPase genes are required for otolith formation and semicircular canal development in zebrafish

Brian Blasiole, Victor A. Canfield, Melissa A. Vollrath, David Huss, Manzoor Ali P.K. Mohideen, J. David Dickman, Keith C. Cheng, Donna M. Fekete, Robert Levenson

Research output: Contribution to journalArticlepeer-review

32 Scopus citations

Abstract

We have investigated the role of Na,K-ATPase genes in zebrafish ear development. Six Na,K-ATPase genes are differentially expressed in the developing zebrafish inner ear. Antisense morpholino knockdown of Na,K-ATPase α1a.1 expression blocked formation of otoliths. This effect was phenocopied by treatment of embryos with ouabain, an inhibitor of Na,K-ATPase activity. The otolith defect produced by morpholinos was rescued by microinjection of zebrafish α1a.1 or rat α1 mRNA, while the ouabain-induced defect was rescued by expression of ouabain-resistant zebrafish α1a.1 or rat α1 mRNA. Knockdown of a second zebrafish α subunit, α1a.2, disrupted development of the semicircular canals. Knockdown of Na,K-ATPase β2b expression also caused an otolith defect, suggesting that the β2b subunit partners with the α1a.1 subunit to form a Na,K-ATPase required for otolith formation. These results reveal novel roles for Na,K-ATPase genes in vestibular system development and indicate that different isoforms play distinct functional roles in formation of inner ear structures. Our results highlight zebrafish gene knockdown-mRNA rescue as an approach that can be used to dissect the functional properties of zebrafish and mammalian Na,K-ATPase genes.

Original languageEnglish (US)
Pages (from-to)148-160
Number of pages13
JournalDevelopmental biology
Volume294
Issue number1
DOIs
StatePublished - Jun 1 2006

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Developmental Biology
  • Cell Biology

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