Characterization of the Transcriptome of Nascent Hair Cells and Identification of Direct Targets of the Atoh1 Transcription Factor
Characterization of the Transcriptome of Nascent Hair Cells and Identification of Direct Targets of the Atoh1 Transcription Factor
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DOI：
10.1523/jneurosci.5083-14.2015
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发表时间：
2015-04-08
影响因子：
5.3
通讯作者：
Groves, Andrew K.
中科院分区：
文献类型：
--
作者：
Cai, Tiantian;Jen, Hsin-I;Groves, Andrew K.
毛细胞是脊椎动物听觉和前庭系统的感觉受体。转录因子Atoh 1对于毛细胞的分化是必要的和足够的，并且在非哺乳动物脊椎动物的毛细胞再生过程中强烈上调。为了鉴定参与毛细胞发育和功能的基因，我们对新生小鼠耳蜗中表达Atoh 1的纯化毛细胞进行了RNA-seq分析。我们在耳蜗毛细胞中鉴定了>600种富集的转录物，其中90%先前未显示在毛细胞中表达。我们根据Atoh 1结合位点在其调控区的存在，并通过分析来自小脑和小肠的Atoh 1 ChIP-seq数据集，确定了233个毛细胞基因作为Atoh 1直接调控的候选基因。我们通过ChIP-PCR证实了其中10个基因是耳蜗中Atoh 1的直接靶点。候选Atoh 1靶基因的鉴定是确定毛细胞发育的基因调控网络的第一步，并可能为将来研究Atoh 1在哺乳动物毛细胞再生中的潜在作用提供信息。
Hair cells are sensory receptors for the auditory and vestibular system in vertebrates. The transcription factor Atoh1 is both necessary and sufficient for the differentiation of hair cells, and is strongly upregulated during hair-cell regeneration in nonmammalian vertebrates. To identify genes involved in hair cell development and function, we performed RNA-seq profiling of purified Atoh1-expressing hair cells from the neonatal mouse cochlea. We identified >600 enriched transcripts in cochlear hair cells, of which 90% have not been previously shown to be expressed in hair cells. We identified 233 of these hair cell genes as candidates to be directly regulated by Atoh1 based on the presence of Atoh1 binding sites in their regulatory regions and by analyzing Atoh1 ChIP-seq datasets from the cerebellum and small intestine. We confirmed 10 of these genes as being direct Atoh1 targets in the cochlea by ChIP-PCR. The identification of candidate Atoh1 target genes is a first step in identifying gene regulatory networks for hair-cell development and may inform future studies on the potential role of Atoh1 in mammalian hair cell regeneration.