Atoh1 expression levels define the fate of rat cochlear nonsensory epithelial cells in vitro.
Atoh1 expression levels define the fate of rat cochlear nonsensory epithelial cells in vitro.
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Atoh1表达水平决定大鼠耳蜗非感觉上皮细胞的体外命运
DOI：
10.3892/mmr.2014.2202
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发表时间：
2014-07
影响因子：
3.4
通讯作者：
Chi FL
中科院分区：
文献类型：
--
作者：
Luo WW;Yang JM;Han Z;Yuan YS;Sheng HB;Liu X;Chi FL
无调性同源物1 （Atoh1）是一种基本的螺旋-环-螺旋转录因子，对内耳毛细胞分化至关重要。先前的研究报道了Atoh1基因转移诱导异位毛细胞样细胞（ehclc）的产生。本研究采用编码Atoh1和报告基因EGFP的人腺病毒血清5型（Ad5）载体，研究了不同Atoh1表达水平和EHCLC形成时间对耳蜗小上皮脊（LER）的影响。在人工耳蜗中加入不同滴度的Ad5-EGFP-atoh1/Ad5-EGFP病毒，并在不同时间点在耳蜗内耳中检测ehclc。结果表明，单独使用GFP不能诱导ehclc。相比之下，在EGFP-atoh1感染LER后2.5-5天，Atoh1表达诱导ehclc，并且随着病毒滴度的不同，ehclc的数量随着时间的推移而增加。较高的Ad5-EGFP-atoh1滴度诱导Atoh1表达增强，导致ehclc增加。较低的Ad5-EGFP-atoh1滴度需要更长的时间来形成EHCLC，而极低的Ad5-EGFP-atoh1滴度仅诱导弱的Atoh1表达，不会触发EHCLC的形成。综上所述，本研究利用适当的Ad5-EGFP-atoh1滴度范围诱导Atoh1表达并随后产生ehclc。结果显示，Atoh1的表达水平决定了LER细胞的命运是ehclc还是非感觉上皮细胞。这一证据可能为今后耳聋基因治疗策略的研究提供重要的指导。
Atonal homolog 1 (Atoh1) is a basic helix-loop-helix transcription factor that is essential for inner ear hair cell differentiation. Previous studies have reported that Atoh1 gene transfer induces the production of ectopic hair cell-like cells (EHCLCs). In the present study, the effect of different Atoh1 expression levels and the duration of EHCLC formation on the lesser epithelial ridge (LER) of cochleae was examined using a human adenovirus serotype 5 (Ad5) vector encoding atoh1 and the reporter gene EGFP. Different Ad5-EGFP-atoh1/Ad5-EGFP virus titers were added to cultured cochlear explants and EHCLCs were detected in the LER at various time points. The results demonstrated that GFP alone did not induce EHCLCs. By contrast, Atoh1 expression induced EHCLCs as early as 2.5–5 days following EGFP-atoh1 infection in the LER and depending upon the viral titer, the number of EHCLCs increased with time. Higher Ad5-EGFP-atoh1 titers induced enhanced Atoh1 expression, resulting in an increase in EHCLCs. Lower Ad5-EGFP-atoh1 titers required more time for EHCLC formation and very low titers of Ad5-EGFP-atoh1 induced only weak Atoh1 expression and did not trigger EHCLC formation. In conclusion, the present study utilized an appropriate Ad5-EGFP-atoh1 titer range to induce Atoh1 expression and the subsequent production of EHCLCs. The results revealed that the Atoh1 expression level defined the fate of LER cells as either EHCLCs or nonsensory epithelial cells. This evidence may provide an important guideline for future studies into gene therapy strategies for the treatment of deafness.
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影响因子：
25
作者：
Zheng, JL;Gao, WQ
通讯作者：
Gao, WQ
影响因子：
64.8
作者：
Gubbels, Samuel P.;Woessner, David W.;Mitchell, John C.;Ricci, Anthony J.;Brigande, John V.
通讯作者：
Brigande, John V.
影响因子：
56.9
作者：
RYALS, BM;RUBEL, EW
通讯作者：
RUBEL, EW
影响因子：
56.9
作者：
Bermingham, NA;Hassan, BA;Zoghbi, HY
通讯作者：
Zoghbi, HY
影响因子：
56.9
作者：
CORWIN, JT;COTANCHE, DA
通讯作者：
COTANCHE, DA