Separate Na,K-ATPase genes are required for otolith formation and semicircular canal development in zebrafish
Separate Na,K-ATPase genes are required for otolith formation and semicircular canal development in zebrafish
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DOI：
10.1016/j.ydbio.2006.02.034
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发表时间：
2006-06-01
影响因子：
2.7
通讯作者：
Levenson, Robert
中科院分区：
文献类型：
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作者：
Blasiole, Brian;Canfield, Victor A.;Levenson, Robert
我们研究了 Na,K-ATPase 基因在斑马鱼耳朵发育中的作用。六个 Na,K-ATP 酶基因在发育中的斑马鱼内耳中差异表达。 Na,K-ATPase α 1a.1 表达的反义吗啉敲低可阻止耳石的形成。通过用哇巴因（一种 Na,K-ATP 酶活性抑制剂）处理胚胎来模拟这种效应。由morpholmos产生的耳石缺损通过显微注射斑马鱼α1a.1或大鼠α1 mRNA来挽救，而哇巴因诱导的缺损则通过表达耐哇巴因的斑马鱼α1a.1或大鼠α1 mRNA来挽救。第二个斑马鱼a亚基α1a.2的敲低破坏了半规管的发育。 Na,K-ATPase beta 2b 表达的敲低也会导致耳石缺陷，表明 beta 2b 亚基与 alpha 1a.1 亚基合作形成耳石形成所需的 Na,K-ATPase。这些结果揭示了Na,K-ATP酶基因在前庭系统发育中的新作用，并表明不同亚型在内耳结构的形成中发挥不同的功能作用。我们的结果强调了斑马鱼基因敲低 mRNA 救援作为一种可用于剖析斑马鱼和哺乳动物 Na,K-ATP 酶基因功能特性的方法。 (c) 2006 Elsevier Inc. 保留所有权利。
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 alpha 1a.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 morpholmos was rescued by microinjection of zebrafish alpha 1a.1 or rat alpha 1 mRNA, while the ouabain-induced defect was rescued by expression of ouabain-resistant zebrafish alpha 1a.1 or rat alpha 1 mRNA. Knockdown of a second zebrafish a subunit, alpha 1a.2, disrupted development of the semicircular canals. Knockdown of Na,K-ATPase beta 2b expression also caused an otolith defect, suggesting that the beta 2b subunit partners with the alpha 1a.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. (c) 2006 Elsevier Inc. All rights reserved.