Global Analysis of Protein Expression of Inner Ear Hair Cells
Global Analysis of Protein Expression of Inner Ear Hair Cells
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DOI：
10.1523/jneurosci.2267-16.2016
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发表时间：
2017-02-01
影响因子：
5.3
通讯作者：
Savas, Jeffrey N.
中科院分区：
文献类型：
--
作者：
Hickox, Ann E.;Wong, Ann C. Y.;Savas, Jeffrey N.
哺乳动物内耳(IE)通过高度特化的细胞和蛋白质来调节听觉和前庭感觉。感觉受体毛细胞(HCs)是利用一系列已知和未知的蛋白质机制传递机械输入和刺激感觉神经元所必需的。为了了解这些独特的有丝分裂后细胞的蛋白质组成，其中不可逆转的蛋白质降解或损伤可能导致听力和平衡受损，我们用串联质谱学分析IE样本，以产生关于蛋白质鉴定和丰度的无偏见的鸟枪式蛋白质组学观点。利用Pou4f3/EGFP转基因小鼠，在Pou4f3驱动的HCS中表达GFP，FACS纯化了一组HC，并将其与来自感觉上皮和整个IE的其他IE亚组进行了分析和比较。我们知道哺乳动物的HC蛋白质组由数百种独特或高表达的蛋白质组成。我们对纯化的HCS进行的全球蛋白质组分析扩展了现有的HC转录组，揭示了以前未检测到的基因产物和异构体特异的蛋白质表达。将我们的蛋白质组数据与小鼠和人类的遗传性听觉/前庭损伤数据库进行比较，证实了HC蛋白质组对正常IE功能的关键作用，为新的、重要的HC基因提供了一个特定于细胞的候选库。通过蛋白质组学鉴定并经免疫组织化学验证的几种蛋白质定位于人类遗传性耳聋基因座，可能代表新的耳聋基因。
The mammalian inner ear (IE) subserves auditory and vestibular sensations via highly specialized cells and proteins. Sensory receptor hair cells (HCs) are necessary for transducing mechanical inputs and stimulating sensory neurons by using a host of known and as yet unknown protein machinery. To understand the protein composition of these unique postmitotic cells, in which irreversible protein degradation or damage can lead to impaired hearing and balance, we analyzed IE samples by tandem mass spectrometry to generate an unbiased, shotgun-proteomics view of protein identities and abundances. By using Pou4f3/eGFP-transgenic mice in which HCs express GFP driven by Pou4f3, we FACS purified a population of HCs to analyze and compare the HC proteome with other IE subproteomes from sensory epithelia and whole IE. Weshow that the mammalian HC proteome comprises hundreds of uniquely or highly expressed proteins. Our global proteomic analysis of purified HCs extends the existing HC transcriptome, revealing previously undetected gene products and isoform-specific protein expression. Comparison of our proteomic data with mouse and human databases of genetic auditory/vestibular impairments confirms the critical role of the HC proteome for normal IE function, providing a cell-specific pool of candidates for novel, important HC genes. Several proteins identified exclusively in HCs by proteomics and verified by immunohistochemistry map to human genetic deafness loci, potentially representing new deafness genes.