An RNA interference-based screen of transcription factor genes identifies pathways necessary for sensory regeneration in the avian inner ear.
An RNA interference-based screen of transcription factor genes identifies pathways necessary for sensory regeneration in the avian inner ear.
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DOI：
10.1523/jneurosci.5456-10.2011
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发表时间：
2011-03-23
期刊：
影响因子：
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通讯作者：
Lovett M
中科院分区：
文献类型：
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作者：
Alvarado DM;Hawkins RD;Bashiardes S;Veile RA;Ku YC;Powder KE;Spriggs MK;Speck JD;Warchol ME;Lovett M
内耳的感觉毛细胞是声音和头部运动的机电换能器。在哺乳动物中，感觉毛细胞的损伤会导致听力或平衡障碍。非哺乳类脊椎动物，如鸟类，在受伤后可以再生毛细胞。在之前的研究中，我们描述了鸡毛细胞再生过程中转录因子基因的表达。在这些研究中，使用激光微束或耳毒性抗生素来损伤感觉上皮（SE）。目前的研究集中在27个基因上，这些基因在再生SE中与先前的研究中未治疗的SE相比上调。这些基因被siRNA敲除，以确定它们对支持细胞增殖的需求，并测量在更大的基因表达网络中产生的变化。我们确定了11个增殖所必需的基因，并确定了其中许多基因之间新的相互作用关系。WNT、PAX和AP1通路的特定组分被证明是支持细胞增殖所必需的。这些通路在WNT4上相交，WNT4也是增殖所必需的。在所需基因中，CCAAT增强子结合蛋白CEBPG在AP1通路中作用于Jun激酶和JUND的下游。WNT共受体LRP5与转录因子BTAF1一样作用于CEBPG的下游。这两种基因也是支持细胞增殖所必需的。这是该类型的首次大规模筛选，表明在内耳毛细胞再生过程中，AP1通路、PAX通路和WNT信号通路在调控支持细胞增殖中的重要交叉。
Sensory hair cells of the inner ear are the mechano-electric transducers of sound and head motion. In mammals, damage to sensory hair cells leads to hearing or balance deficits. Non-mammalian vertebrates such as birds can regenerate hair cells after injury. In a previous study, we characterized transcription factor gene expression during chicken hair cell regeneration. In those studies, a laser micro-beam or ototoxic antibiotics were used to damage the sensory epithelia (SE). The current study focused on 27 genes that were up-regulated in regenerating SE compared to untreated SE in the previous study. Those genes were knocked down by siRNA, to determine their requirement for supporting cell proliferation and to measure resulting changes in the larger network of gene expression. We identified 11 genes necessary for proliferation and also identified novel interactive relationships between many of them. Defined components of the WNT, PAX and AP1 pathways were shown to be required for supporting cell proliferation. These pathways intersect on WNT4, which is also necessary for proliferation. Among the required genes, the CCAAT enhancer binding protein, CEBPG, acts downstream of Jun Kinase and JUND in the AP1 pathway. The WNT co-receptor LRP5 acts downstream of CEBPG as does the transcription factor BTAF1. Both of these genes are also necessary for supporting cell proliferation. This is the first large scale screen of its type and suggests an important intersection between the AP1 pathway, the PAX pathway and WNT signaling in the regulation of supporting cell proliferation during inner ear hair cell regeneration.