Lingo-1 shRNA and Notch signaling inhibitor DAPT promote differentiation of neural stem/progenitor cells into neurons
Lingo-1 shRNA and Notch signaling inhibitor DAPT promote differentiation of neural stem/progenitor cells into neurons
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Lingo-1 shRNA 和 Notch 信号抑制剂 DAPT 促进神经干/祖细胞分化为神经元
DOI：
10.1016/j.brainres.2015.11.029
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发表时间：
2016-03-01
期刊：
影响因子：
2.9
通讯作者：
Yang, Ruirui
中科院分区：
文献类型：
--
作者：
Wang, Jue;Ye, Zhizhong;Yang, Ruirui
确定调节神经干/祖细胞分化为神经元、少突胶质细胞和星形胶质细胞的外源因子是脊髓损伤（SCI）临床治疗的重要一步。 Notch 通路抑制神经干/祖细胞的分化，Lingo-1 是髓鞘形成和轴突生长的强负调节因子。虽然 Lingo-1 shRNA 和 Notch 通路抑制剂 N-[N-(3, 5-二氟苯乙酰基)1-丙氨酰]-S-苯基甘氨酸丁酯 (DAPT) 分别用于帮助修复 SCI，但结果并不令人满意。在这里，我们研究并阐明了 Lingo-1 shRNA 和 DAPT 对神经干/祖细胞分化影响的初步机制。我们发现来自E14大鼠胚胎的神经干/祖细胞表达Nestin、Sox-2和Lingo-1，并且我们使用编码Lingo-1 shRNA的慢病毒载体优化了神经干/祖细胞的转导。 DAPT 的添加降低了 Notch 胞内结构域 (NICD) 以及下游基因 Hest 和 Hes5 的表达。 DAPT 处理的细胞中 NeuN、CNPase 和 GFAP 的表达以及 Lingo-1 shRNA 处理的细胞中 NeuN 的表达证实了神经干/祖细胞分化为神经元、少突胶质细胞和星形胶质细胞。这些结果表明，虽然Lingo-1 shRNA和Notch信号抑制剂DAPT都促进神经干细胞分化为神经元，但只有DAPT能够驱动神经干/祖细胞分化为少突胶质细胞和星形胶质细胞。由于我们能够证明 Lingo-1 shRNA 和 DAPT 都可以驱动神经干/祖细胞分化，因此我们的数据可能有助于使用 Lingo-1 shRNA 和 DAPT 开发更有效的 SCI 疗法。 (C) 2015 Elsevier B.V. 保留所有权利。
Determination of the exogenous factors that regulate differentiation of neural stem/progenitor cells into neurons, oligodendrocytes and astrocytes is an important step in the clinical therapy of spinal cord injury (SCI). The Notch pathway inhibits the differentiation of neural stem/progenitor cells and Lingo-1 is a strong negative regulator for myelination and axon growth. While Lingo-1 shRNA and N-[N-(3, 5-difluorophenacetyl)1-alanyl]-S-Phenylglycinet-butylester (DAPT), a Notch pathway inhibitor, have been used separately to help repair SCI, the results have been unsatisfactory. Here we investigated and elucidated the preliminary mechanism for the effect of Lingo-1 shRNA and DAPT on neural stem/progenitor cells differentiation. We found that neural stem/progenitor cells from E14 rat embryos expressed Nestin, Sox-2 and Lingo-1, and we optimized the transduction of neural stem/progenitor cells using lentiviral vectors encoding Lingo-1 shRNA. The addition of DAPT decreased the expression of Notch intracellular domain (NICD) as well as the downstream genes Hest and Hes5. Expression of NeuN, CNPase and GFAP in DAPT treated cells and expression of NeuN in Lingo-1 shRNA treated cells confirmed differentiation of neural stem/progenitor cells into neurons, oligodendrocytes and astrocytes. These results revealed that while Lingo-1 shRNA and Notch signaling inhibitor DAPT both promoted differentiation of neural stem cells into neurons, only DAPT was capable of driving neural stem/progenitor cells differentiation into oligodendrocytes and astrocytes. Since we were able to show that both Lingo-1 shRNA and DAPT could drive neural stem/progenitor cells differentiation, our data might aid the development of more effective SCI therapies using Lingo-1 shRNA and DAPT. (C) 2015 Elsevier B.V. All rights reserved.