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ZIRC
ZFIN ID: ZDB-PUB-060403-5
Separate Na,K-ATPase genes are required for otolith formation and semicircular canal development in zebrafish
Blasiole, B., Canfield, V.A., Vollrath, M.A., Huss, D., Mohideen, M.A., Dickman, J.D., Cheng, K.C., Fekete, D.M., and Levenson, R.
Date: 2006
Source: Developmental Biology   294(1): 148-160 (Journal)
Registered Authors: Blasiole, Brian, Canfield, Victor, Cheng, Keith C., Fekete, Donna Marie, Levenson, Robert, Mohideen, Manzoor Pallithotangal
Keywords: Zebrafish, Na,K-ATPase, Sodium pump, Morpholino, Inner ear, Otolith, Semicircular canal, mRNA rescue
MeSH Terms:
  • Animals
  • Ear/growth & development
  • Embryo, Nonmammalian
  • Isoenzymes
  • Morphogenesis
  • Otolithic Membrane/enzymology*
  • Protein Subunits
  • Rats
  • Semicircular Canals/enzymology*
  • Sodium-Potassium-Exchanging ATPase/genetics*
  • Sodium-Potassium-Exchanging ATPase/physiology
  • Zebrafish
PubMed: 16566913 Full text @ Dev. Biol.
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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 alpha1a.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 alpha1a.1 or rat alpha1 mRNA, while the ouabain-induced defect was rescued by expression of ouabain-resistant zebrafish alpha1a.1 or rat alpha1 mRNA. Knockdown of a second zebrafish alpha subunit, alpha1a.2, disrupted development of the semicircular canals. Knockdown of Na,K-ATPase beta2b expression also caused an otolith defect, suggesting that the beta2b subunit partners with the alpha1a.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.
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