Scheffer Déborah I, Shen Jun, Corey David P, Chen Zheng-Yi
Department of Neurobiology and Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, Department of Otology and Laryngology, Harvard Medical School and Massachusetts Eye and Ear, Boston, Massachusetts 02114, and.
Department of Neurobiology and Howard Hughes Medical Institute, Harvard Medical School, Boston, Massachusetts 02115, Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115.
J Neurosci. 2015 Apr 22;35(16):6366-80. doi: 10.1523/JNEUROSCI.5126-14.2015.
Hair cells of the inner ear are essential for hearing and balance. As a consequence, pathogenic variants in genes specifically expressed in hair cells often cause hereditary deafness. Hair cells are few in number and not easily isolated from the adjacent supporting cells, so the biochemistry and molecular biology of hair cells can be difficult to study. To study gene expression in hair cells, we developed a protocol for hair cell isolation by FACS. With nearly pure hair cells and surrounding cells, from cochlea and utricle and from E16 to P7, we performed a comprehensive cell type-specific RNA-Seq study of gene expression during mouse inner ear development. Expression profiling revealed new hair cell genes with distinct expression patterns: some are specific for vestibular hair cells, others for cochlear hair cells, and some are expressed just before or after maturation of mechanosensitivity. We found that many of the known hereditary deafness genes are much more highly expressed in hair cells than surrounding cells, suggesting that genes preferentially expressed in hair cells are good candidates for unknown deafness genes.
内耳的毛细胞对听力和平衡至关重要。因此，在毛细胞中特异性表达的基因中的致病变异通常会导致遗传性耳聋。毛细胞数量稀少，且不易与相邻的支持细胞分离，因此毛细胞的生物化学和分子生物学研究可能具有挑战性。为了研究毛细胞中的基因表达，我们开发了一种通过荧光激活细胞分选（FACS）分离毛细胞的方案。利用从耳蜗和椭圆囊、从胚胎第16天到出生后第7天获得的近乎纯的毛细胞及周围细胞，我们对小鼠内耳发育过程中的基因表达进行了全面的细胞类型特异性RNA测序研究。表达谱分析揭示了具有不同表达模式的新的毛细胞基因：一些对前庭毛细胞具有特异性，另一些对耳蜗毛细胞具有特异性，还有一些在机械敏感性成熟之前或之后表达。我们发现，许多已知的遗传性耳聋基因在毛细胞中的表达比周围细胞高得多，这表明在毛细胞中优先表达的基因是未知耳聋基因的良好候选者。
