Developmental and Functional Hair Cell-Like Cells Induced by Atoh1 Overexpression in the Adult Mammalian Cochlea In Vitro.
Developmental and Functional Hair Cell-Like Cells Induced by Atoh1 Overexpression in the Adult Mammalian Cochlea In Vitro.
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DOI：
10.1155/2020/8885813
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发表时间：
2020
影响因子：
3.1
通讯作者：
Yang J
中科院分区：
文献类型：
--
作者：
Kong L;Xin Y;Chi F;Chen J;Yang J
哺乳动物耳蜗中的毛细胞一旦受损就不能自发再生，导致永久性听力损失。已有研究表明，Atoh1过表达可诱导新生啮齿动物耳蜗毛细胞样细胞(HcLCs)的形成，但在成年哺乳动物中难以实现。在这项研究中，我们使用三维耳蜗培养系统和腺病毒介导的递送载体在成年小鼠耳蜗中过表达Atoh1。病毒感染后3d(3DVI)即可在体外成功诱导出HCLCs，且数量随时间增加而增加。在培养环境中，HCLC呈myosin7a阳性，可与残留的HCs区分开来。同时，膜片钳结果显示，人肝细胞可记录到非激活外向钾电流(持续外向钾电流)，其幅度随时间延长而增大，与正常肝细胞相似。此外，在一些HCLC中记录到了瞬时的HCN电流，这表明这些HCLC经历了一个与正常HC相似的发育阶段。我们还比较了成年小鼠和天然的小鼠肝细胞的电生理特征，发现肝细胞逐渐成熟，与正常的肝细胞相似。同时，成年小鼠的HCLC与发育期的HC具有相同的束状结构。然而，即使在13DVI时，这些HCLC也不表达prestin，这是外毛细胞的一个特殊标记。这些结果表明，Atoh1过表达可诱导成年哺乳动物耳蜗内HCLC的形成，并且这些HCLC具有功能，并且经历了与正常HCS相似的发育过程。
Hair cells (HCs) in the mammalian cochleae cannot spontaneously regenerate once damaged, resulting in permanent hearing loss. It has been shown that Atoh1 overexpression induces hair cell-like cells (HCLCs) in the cochlea of newborn rodents, but this is hard to achieve in adult mammals. In this study, we used a three-dimensional cochlear culture system and an adenoviral-mediated delivery vector to overexpress Atoh1 in adult mouse cochleae. HCLCs were successfully induced from 3 days after virus infection (3 DVI) in vitro, and the number increased with time. HCLCs were myosin7a positive and distinguishable from remnant HCs in a culture environment. Meanwhile, patch-clamp results showed that noninactive outward potassium currents (sustained outward potassium currents) could be recorded in HCLCs and that their magnitude increased with time, similar to normal HCs. Furthermore, transient HCN currents were recorded in some HCLCs, indicating that the HCLCs experienced a developmental stage similar to normal HCs. We also compared the electrophysiological features of HCLCs from adult mice with native HCs and found the HCLCs gradually matured, similar to the normal HCs. Meanwhile, HCLCs from adult mice possessed the same bundles as developmental HCs. However, these HCLCs did not express prestin, which is a special marker for outer hair cells (OHCs), even at 13 DVI. These results demonstrate that Atoh1 overexpression induces HCLC formation in the adult mammalian cochlea and that these HCLCs were functional and experienced a developmental process similar to that of normal HCs.
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影响因子：
2.8
作者：
Kawamoto K;Izumikawa M;Beyer LA;Atkin GM;Raphael Y
通讯作者：
Raphael Y
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Brignull HR;Raible DW;Stone JS
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Stone JS
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2.5
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通讯作者：
Stone, Jennifer S.
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4.5
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56.9
作者：
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通讯作者：
NEVILL, G