Knocking down of heat-shock protein 27 directs differentiation of functional glutamatergic neurons from placenta-derived multipotent cells.
Knocking down of heat-shock protein 27 directs differentiation of functional glutamatergic neurons from placenta-derived multipotent cells.
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DOI：
10.1038/srep30314
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发表时间：
2016-07-22
影响因子：
4.6
通讯作者：
Chien CC
中科院分区：
文献类型：
--
作者：
Cheng YC;Huang CJ;Lee YJ;Tien LT;Ku WC;Chien R;Lee FK;Chien CC
这项研究介绍了人胎盘来源的多能细胞(PDMCs)作为一个来源，功能谷氨酸能神经元可以从中衍生。我们发现，在神经元分化过程中，小分子热休克蛋白27(HSP27)表达下调。在小鼠胚胎发育过程中，HSP27表达的时间和空间分布与神经元蛋白呈倒置分布。HSP27在干细胞中的过表达导致神经元分化停止；然而，HSP27的敲除使PDMCs分化为神经元的能力显著增强。这些神经元形成突触网络，多个神经元标记呈阳性染色。此外，细胞现象，包括在HSP27沉默的PDMCs中没有凋亡和罕见的增殖，结合分子事件，如切割的caspase-3和切割的Nanog失去茎，表明HSP27位于神经元分化的上游，并限制了这一过程。此外，谷氨酸处理后诱导的神经元细胞内钙离子浓度升高。这些分化的细胞共表达N-甲基-D-天冬氨酸受体、囊泡谷氨酸转运体和突触体相关蛋白25，但不表达酪氨酸羟化酶、胆碱乙酰转移酶或谷氨酸脱羧酶67。因此，我们得出结论：HSP27沉默的PDMCs分化为具有功能谷氨酸能神经元特征的神经元。
This study presents human placenta-derived multipotent cells (PDMCs) as a source from which functional glutamatergic neurons can be derived. We found that the small heat-shock protein 27 (HSP27) was downregulated during the neuronal differentiation process. The in vivo temporal and spatial profiles of HSP27 expression were determined and showed inverted distributions with neuronal proteins during mouse embryonic development. Overexpression of HSP27 in stem cells led to the arrest of neuronal differentiation; however, the knockdown of HSP27 yielded a substantially enhanced ability of PDMCs to differentiate into neurons. These neurons formed synaptic networks and showed positive staining for multiple neuronal markers. Additionally, cellular phenomena including the absence of apoptosis and rare proliferation in HSP27-silenced PDMCs, combined with molecular events such as cleaved caspase-3 and the loss of stemness with cleaved Nanog, indicated that HSP27 is located upstream of neuronal differentiation and constrains that process. Furthermore, the induced neurons showed increasing intracellular calcium concentrations upon glutamate treatment. These differentiated cells co-expressed the N-methyl-D-aspartate receptor, vesicular glutamate transporter, and synaptosomal-associated protein 25 but did not show expression of tyrosine hydroxylase, choline acetyltransferase or glutamate decarboxylase 67. Therefore, we concluded that HSP27-silenced PDMCs differentiated into neurons possessing the characteristics of functional glutamatergic neurons.