Tang Feng, Chen Xiaoling, Jia Lifeng, Li Hai, Li Jingya, Yuan Wei
Department of Otolaryngology, Southwest Hospital, Third Military Medical University, Chongqing, China.
Front Mol Neurosci. 2020 Jan 21;12:332. doi: 10.3389/fnmol.2019.00332. eCollection 2019.
Tonotopic differences in the structure and physiological function, e.g., synapse number, membrane properties, Ca channels, Ca dependence of exocytosis and vesicle pool replenishment of inner hair cells (IHCs) along the longitudinal cochlear axis have recently been discovered, suggesting different gene expression patterns of IHCs. To determine whether IHCs present different gene expression patterns along the longitudinal cochlear axis, apical and basal IHCs were collected separately using the suction pipette technique from adult mouse cochleae for RNA-seq and genome-wide transcriptome analysis. We found 689 annotated genes showed more than 2-fold increase in expression. Interestingly, 93.4% of the differentially expressed genes (DEGs) was upregulated in apical IHCs. Although a subset of genes that related to IHC machinery and deafness were found to be differentially expressed, a gradient of gene expression was indeed detected in , and , which may play putative roles in the Ca buffering and survival regulation. The expression of these genes was validated by real-time quantitative PCR (RT-qPCR) or immunostaining. We conclude that IHCs from different mouse cochlear longitudinal position have different gene expression profiles. Our data might serve as a valuable resource for exploring the molecular mechanisms underlying different biological properties as well as the survival regulation of IHCs.
最近发现,沿着耳蜗纵轴,内毛细胞(IHC)在结构和生理功能方面存在音调定位差异,例如突触数量、膜特性、钙通道、胞吐作用的钙依赖性和囊泡池补充等,这表明IHC存在不同的基因表达模式。为了确定IHC沿耳蜗纵轴是否呈现不同的基因表达模式,使用吸管技术从成年小鼠耳蜗中分别收集顶端和基部的IHC,用于RNA测序和全基因组转录组分析。我们发现689个注释基因的表达增加了两倍以上。有趣的是,93.4%的差异表达基因(DEG)在顶端IHC中上调。尽管发现与IHC机制和耳聋相关的一部分基因存在差异表达,但确实在[具体物质或区域未明确给出]中检测到了基因表达梯度,这可能在钙缓冲和存活调节中发挥假定作用。这些基因的表达通过实时定量PCR(RT-qPCR)或免疫染色进行了验证。我们得出结论,来自不同小鼠耳蜗纵向位置的IHC具有不同的基因表达谱。我们的数据可能为探索IHC不同生物学特性以及存活调节的分子机制提供有价值的资源。