Actomyosin contractility plays a role in MAP2 expression during nanotopography-directed neuronal differentiation of human embryonic stem cells
Actomyosin contractility plays a role in MAP2 expression during nanotopography-directed neuronal differentiation of human embryonic stem cells
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DOI：
10.1016/j.biomaterials.2015.01.003
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发表时间：
2015-04-01
期刊：
影响因子：
14
通讯作者：
Yim, Evelyn K. F.
中科院分区：
文献类型：
--
作者：
Ankam, Soneela;Lim, Choon Kiat;Yim, Evelyn K. F.
多能人胚胎干细胞（hESCs）具有根据特定环境因素分化为不同谱系的能力。我们之前已经证明hESCs可以分化为神经元或神经胶质细胞，这取决于其底物地形的排列、几何形状和大小。特别是，各向异性图案的底物，如光栅，被发现有利于hESCs分化为神经元而不是胶质细胞。在这项研究中，我们的目的是阐明地形诱导hESCs向神经元谱系分化的潜在机制。我们发现，由纳米光栅地形诱导的高肌动球蛋白收缩性对神经元成熟至关重要。用肌球蛋白II抑制剂（blebbistatin）和肌球蛋白轻链激酶抑制剂（ML-7）处理细胞可大大降低微管相关蛋白2 （MAP2）的表达水平。另一方面，我们的qPCR阵列结果显示，在纳米光栅底物上生长的hESCs中，PAX5、BRN3A和NEUROD1的表达高于无图案底物，这表明这些基因可能参与了地形介导的hESCs神经元分化。有趣的是，YAP定位于分化hESCs的细胞质中。综上所述，我们的研究为理解hESCs神经元分化过程中地形线索的机械转导提供了新的见解。(C) 2015 Elsevier Ltd.版权所有。
Pluripotent human embryonic stem cells (hESCs) have the capability of differentiating into different lineages based on specific environmental cues. We had previously shown that hESCs can be primed to differentiate into either neurons or glial cells, depending on the arrangement, geometry and size of their substrate topography. In particular, anisotropically patterned substrates like gratings were found to favour the differentiation of hESCs into neurons rather than glial cells. In this study, our aim is to elucidate the underlying mechanisms of topography-induced differentiation of hESCs towards neuronal lineages. We show that high actomyosin contractility induced by a nano-grating topography is crucial for neuronal maturation. Treatment of cells with the myosin II inhibitor (blebbistatin) and myosin light chain kinase inhibitor (ML-7) greatly reduces the expression level of microtubule-associated protein 2 (MAP2). On the other hand, our qPCR array results showed that PAX5, BRN3A and NEUROD1 were highly expressed in hESCs grown on nano-grating substrates as compared to unpatterned substrates, suggesting the possible involvement of these genes in topography-mediated neuronal differentiation of hESCs. Interestingly, YAP was localized to the cytoplasm of differentiating hESCs. Taken together, our study has provided new insights in understanding the mechanotransduction of topographical cues during neuronal differentiation of hESCs. (C) 2015 Elsevier Ltd. All rights reserved.