Seiler Christoph, Ben-David Orit, Sidi Samuel, Hendrich Oliver, Rusch Alfons, Burnside Beth, Avraham Karen B, Nicolson Teresa
Max-Planck-Institut fur Entwicklungsbiologie, 72076 Tubingen, Germany.
Dev Biol. 2004 Aug 15;272(2):328-38. doi: 10.1016/j.ydbio.2004.05.004.
Unconventional myosins have been associated with hearing loss in humans, mice, and zebrafish. Mutations in myosin VI cause both recessive and dominant forms of nonsyndromic deafness in humans and deafness in Snell's waltzer mice associated with abnormal fusion of hair cell stereocilia. Although myosin VI has been implicated in diverse cellular processes such as vesicle trafficking and epithelial morphogenesis, the role of this protein in the sensory hair cells remains unclear. To investigate the function of myosin VI in zebrafish, we cloned and examined the expression pattern of myosin VI, which is duplicated in the zebrafish genome. One duplicate, myo6a, is expressed in a ubiquitous pattern during early development and at later stages, and is highly expressed in the brain, gut, and kidney. myo6b, on the other hand, is predominantly expressed in the sensory epithelium of the ear and lateral line at all developmental stages examined. Both molecules have different splice variants expressed in these tissues. Using a candidate gene approach, we show that myo6b is satellite, a gene responsible for auditory/vestibular defects in zebrafish larvae. Examination of hair cells in satellite mutants revealed that stereociliary bundles are irregular and disorganized. At the ultrastructural level, we observed that the apical surface of satellite mutant hair cells abnormally protrudes above the epithelium and the membrane near the base of the stereocilia is raised. At later stages, stereocilia fused together. We conclude that zebrafish myo6b is required for maintaining the integrity of the apical surface of hair cells, suggesting a conserved role for myosin VI in regulation of actin-based interactions with the plasma membrane.
非常规肌球蛋白与人类、小鼠和斑马鱼的听力损失有关。肌球蛋白VI的突变会导致人类隐性和显性非综合征性耳聋,以及斯奈尔华尔兹小鼠的耳聋,这与毛细胞静纤毛的异常融合有关。尽管肌球蛋白VI参与了多种细胞过程,如囊泡运输和上皮形态发生,但该蛋白在感觉毛细胞中的作用仍不清楚。为了研究斑马鱼中肌球蛋白VI的功能,我们克隆并检测了斑马鱼基因组中重复的肌球蛋白VI的表达模式。其中一个副本myo6a在早期发育和后期以普遍存在的模式表达,在脑、肠道和肾脏中高度表达。另一方面,myo6b在所有检测的发育阶段主要在耳朵和侧线的感觉上皮中表达。这两个分子在这些组织中都有不同的剪接变体表达。使用候选基因方法,我们发现myo6b是satellite,一个导致斑马鱼幼虫听觉/前庭缺陷的基因。对satellite突变体中的毛细胞进行检查发现,静纤毛束不规则且紊乱。在超微结构水平上,我们观察到satellite突变体毛细胞的顶端表面异常突出于上皮细胞之上,静纤毛基部附近的膜升高。在后期,静纤毛融合在一起。我们得出结论,斑马鱼myo6b是维持毛细胞顶端表面完整性所必需的,这表明肌球蛋白VI在调节基于肌动蛋白与质膜的相互作用中具有保守作用。