Characterization of Functional Primary Cilia in Human Induced Pluripotent Stem Cell-Derived Neurons
Characterization of Functional Primary Cilia in Human Induced Pluripotent Stem Cell-Derived Neurons
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DOI：
10.1007/s11064-019-02806-4
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发表时间：
2019-04
影响因子：
4.4
通讯作者：
D. Miki;Y. Kobayashi;Tomoya Okada;Tatuso Miyamoto;N. Takei;Y. Sekino;Noriko Koganezawa;T. Shirao;Yumiko Saito
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文献类型：
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作者：
D. Miki;Y. Kobayashi;Tomoya Okada;Tatuso Miyamoto;N. Takei;Y. Sekino;Noriko Koganezawa;T. Shirao;Yumiko Saito
人类诱导多能干细胞的研究进展为生物医学研究和临床应用提供了新的可能。从HiPSCs分化而来的神经元可能是开发各种神经系统疾病的新治疗方法的有前途的工具。然而，HiPSC来源的神经元功能成熟的详细过程仍然知之甚少。在这里，我们分析了hPSC来源的皮质神经元的发育结构，icell GlutaNeurons，重点放在初级纤毛上，初级纤毛是从大多数生长受阻的脊椎动物细胞表面突出的单一感觉细胞器。为了确定神经元纤毛的特征，培养了不同时期的细胞，并用纤毛标记Arl13b和MAP2的抗体进行免疫组织化学染色。原生纤毛在培养天数内即可检测到，且随着培养天数的增加，原生纤毛的发生率和长度增加。用情绪稳定剂锂治疗导致初级纤毛长度延长，而用促食欲素神经肽黑素浓缩激素治疗导致icell谷氨酸神经元纤毛长度缩短。目前的发现表明，icell谷氨酸神经元与调节纤毛长度的信号机制一起发育神经元初级纤毛。我们将初级纤毛作为细胞天线的方法对于评估神经元成熟度和验证HiPSC来源神经元中的药物都是有用的。
Recent advances in human induced pluripotent stem cells (hiPSCs) offer new possibilities for biomedical research and clinical applications. Neurons differentiated from hiPSCs may be promising tools to develop novel treatment methods for various neurological diseases. However, the detailed process underlying functional maturation of hiPSC-derived neurons remains poorly understood. Here, we analyze the developmental architecture of hiPSC-derived cortical neurons, iCell GlutaNeurons, focusing on the primary cilium, a single sensory organelle that protrudes from the surface of most growth-arrested vertebrate cells. To characterize the neuronal cilia, cells were cultured for various periods and evaluated immunohistochemically by co-staining with antibodies against ciliary markers Arl13b and MAP2. Primary cilia were detected in neurons within days, and their prevalence and length increased with increasing days in culture. Treatment with the mood stabilizer lithium led to primary cilia length elongation, while treatment with the orexigenic neuropeptide melanin-concentrating hormone caused cilia length shortening in iCell GlutaNeurons. The present findings suggest that iCell GlutaNeurons develop neuronal primary cilia together with the signaling machinery for regulation of cilia length. Our approach to the primary cilium as a cellular antenna can be useful for both assessment of neuronal maturation and validation of pharmaceutical agents in hiPSC-derived neurons.