Mhatre Anand N, Li Jiang, Kim Yuil, Coling Donald E, Lalwani Anil K
Department of Otolaryngology-Head and Neck Surgery, University of California San Francisco, San Francisco, California, USA.
J Neurosci Res. 2004 May 1;76(3):296-305. doi: 10.1002/jnr.20065.
MYH9 encoding a nonmuscle myosin heavy chain has been linked to nonsyndromic and syndromic forms of autosomal dominant hereditary hearing loss, suggesting a critical biological role of this motor protein in the auditory organ. While Myh9 expression has been described in the adult mouse, critical parameters pertaining to its developmental expression remain to be characterized. The current study describes cloning of the mouse Myh9 cDNA and the temporal onset and spatial distribution of Myh9 expression in the inner ear of the developing fetus, the neonate, and the adult. The cloned Myh9 cDNA contained two single-base-pair differences from the published genomic sequence: T990C (G330G) and T5198A (L1733Q). Immunoblotting of embryonic (E15.5) and adult tissues from several organs, including the cochlea, identified a single 250-kDa anti-Myh9-immunoreactive band, supporting an absence of Myh9 splice variants in the fetus and the adult. In situ expression analysis identified Myh9 distributed within the epithelial layer of the otic vesicle at E10.5. Myh9 expression was found to persist within the epithelia surrounding the cochlear duct at E13.5 and E16.5. The sensory cells of the developing cochlea were positive for Myh9 expression at E16.5. Within the neonate and the adult cochlea, Myh9 expression was observed within the sensory hair cells and the supporting hair cells of the organ of Corti, the spiral ligament, and the spiral limbus, but not in the stria vascularis. Identification of Myh9 in the developing and mature inner ear suggests a role for this protein in the development and maintenance of auditory function.
编码非肌肉肌球蛋白重链的MYH9与常染色体显性遗传性听力损失的非综合征型和综合征型有关,这表明这种运动蛋白在听觉器官中具有关键的生物学作用。虽然在成年小鼠中已经描述了Myh9的表达,但其发育表达的关键参数仍有待确定。当前的研究描述了小鼠Myh9 cDNA的克隆以及Myh9在发育中的胎儿、新生儿和成年小鼠内耳中的表达时间起始和空间分布。克隆的Myh9 cDNA与已发表的基因组序列存在两个单碱基对差异:T990C(G330G)和T5198A(L1733Q)。对包括耳蜗在内的多个器官的胚胎(E15.5)和成年组织进行免疫印迹分析,确定了一条单一的250 kDa抗Myh9免疫反应带,这支持了胎儿和成年小鼠中不存在Myh9剪接变体。原位表达分析确定在E10.5时Myh9分布在耳泡的上皮层内。发现在E13.5和E16.5时,Myh9在耳蜗管周围的上皮中持续表达。在E16.5时,发育中的耳蜗感觉细胞Myh9表达呈阳性。在新生儿和成年耳蜗中,在柯蒂氏器的感觉毛细胞和支持毛细胞、螺旋韧带和螺旋缘中观察到Myh9表达,但在血管纹中未观察到。在发育中和成熟的内耳中鉴定出Myh9表明该蛋白在听觉功能的发育和维持中发挥作用。