TY - JOUR
T1 - Separate Na,K-ATPase genes are required for otolith formation and semicircular canal development in zebrafish
AU - Blasiole, Brian
AU - Canfield, Victor A.
AU - Vollrath, Melissa A.
AU - Huss, David
AU - Mohideen, Manzoor Ali P.K.
AU - Dickman, J. David
AU - Cheng, Keith C.
AU - Fekete, Donna M.
AU - Levenson, Robert
N1 - Funding Information:
The authors are grateful to Inna Hughes for her critical reading of the manuscript, Deb Biesemeier for assistance with histology, and Brian Coon for confocal microscopy. This work was supported by NIH grant MH 068789 and a NARSAD Distinguished Investigator Award to R.L.
PY - 2006/6/1
Y1 - 2006/6/1
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=33646239360&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33646239360&partnerID=8YFLogxK
U2 - 10.1016/j.ydbio.2006.02.034
DO - 10.1016/j.ydbio.2006.02.034
M3 - Article
C2 - 16566913
AN - SCOPUS:33646239360
SN - 0012-1606
VL - 294
SP - 148
EP - 160
JO - Developmental biology
JF - Developmental biology
IS - 1
ER -