Laser-capture micro dissection combined with next-generation sequencing analysis of cell type-specific deafness gene expression in the mouse cochlea
Laser-capture micro dissection combined with next-generation sequencing analysis of cell type-specific deafness gene expression in the mouse cochlea
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DOI：
10.1016/j.heares.2017.02.017
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发表时间：
2017-05-01
期刊：
影响因子：
2.8
通讯作者：
Usami, Shin-ichi
中科院分区：
文献类型：
--
作者：
Nishio, Shin-ya;Takumi, Yutaka;Usami, Shin-ichi
耳蜗植入术（CI）直接刺激耳蜗神经，是重度至极重度感音神经性听力损失患者最有效和最广泛使用的医疗干预措施。推测听力损失的病因学对CI结果具有主要影响，特别是在螺旋神经节区域中优先表达的基因突变导致的病例中。我们进行了激光捕获显微切割结合下一代测序分析，并确定了所有已知耳聋的表达水平，Corti器、螺旋神经节、侧壁和螺旋肢中的相关基因。其结果与以往免疫细胞化学或原位杂交的结果基本一致。作为一个值得注意的结果，与多种综合征性听力损失相关的基因（例如用于Perrault综合征的Clpp、Hars2、Hsdl7b4、Lars2，用于Treacher柯林斯综合征的Poir1c和Polr1d，用于Norrie病的Ndp，用于Kallmann综合征的kal，用于Waardenburg综合征的Edn3和Snail，用于Alport综合征的Col4a3，用于CHARGE综合征的Sema3e，用于Sticker综合征的Col9a1，Cdh 23，Cib 2，Ciml，Pcdh15、Ushlc、Ush2a、Whrn（Usher综合征）和Wfs1（Wolfram综合征）在耳蜗螺旋神经节中的表达水平高于其他部位。(C)2017年，作者。由爱思唯尔公司出版
Cochlear implantation (CI), which directly stimulates the cochlear nerves, is the most effective and widely used medical intervention for patients with severe to profound sensorineural hearing loss. The etiology of the hearing loss is speculated to have a major influence of CI outcomes, particularly in cases resulting from mutations in genes preferentially expressed in the spiral ganglion region.To elucidate precise gene expression levels in each part of the cochlea, we performed laser-capture micro dissection in combination with next-generation sequencing analysis and determined the expression levels of all known deafness-associated genes in the organ of Corti, spiral ganglion, lateral wall, and spiral limbs. The results were generally consistent with previous reports based on immunocytochemistry or in situ hybridization. As a notable result, the genes associated with many kinds of syndromic hearing loss (such as Clpp, Hars2, Hsdl7b4, Lars2 for Perrault syndrome, Poir1c and Polr1d for Treacher Collins syndrome, Ndp for Norrie Disease, kal for Kallmann syndrome, Edn3 and Snail for Waardenburg Syndrome, Col4a3 for Alport syndrome, Sema3e for CHARGE syndrome, Col9a1 for Sticker syndrome, Cdh23, Cib2, Ciml, Pcdh15, Ushlc, Ush2a, Whrn for Usher syndrome and Wfs1 for Wolfram syndrome) showed higher levels of expression in the spiral ganglion than in other parts of the cochlea.This dataset will provide a base for more detailed analysis in order to clarify gene functions in the cochlea as well as predict CI outcomes based on gene expression data. (C) 2017 The Authors. Published by Elsevier B.V.