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Characterization of Functional Primary Cilia in Human Induced Pluripotent Stem Cell-Derived Neurons

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Abstract

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.

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Acknowledgements

The study was supported by research grants (AMED JP17bk0104077 to T.S.) and the Ministry of Education, Culture, Sports, Science and Technology of Japan (KAKENHI 16K07053 to N.T and 15K06775 to Y.S.).

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Author notes

  1. Daisuke Miki and Yuki Kobayashi have contributed equally to this work.

Authors and Affiliations

  1. Graduate School of Integrated Arts and Sciences, Hiroshima University, 1-7-1 Kagamiyama, Higashi-Hiroshima, Hiroshima, 739-8521, Japan

    Daisuke Miki, Yuki Kobayashi, Tomoya Okada & Yumiko Saito

  2. Department of Genetics and Cell Biology, Research Institute for Radiation Biology and Medicine, Hiroshima University, Hiroshima, 734-8553, Japan

    Tatuso Miyamoto

  3. Department of Molecular Neurobiology, Brain Research Institute, Niigata University, Niigata, 951-8585, Japan

    Nobuyuki Takei

  4. Laboratory of Chemical Pharmacology, Graduate School of Pharmaceutical Sciences, University of Tokyo, Tokyo, 113-0033, Japan

    Yuko Sekino

  5. Department of Neurobiology and Behavior, Graduate School of Medicine, Gunma University, Maebashi, 371-8511, Japan

    Noriko Koganezawa & Tomoaki Shirao

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  1. Daisuke Miki
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Correspondence to Yumiko Saito.

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Miki, D., Kobayashi, Y., Okada, T. et al. Characterization of Functional Primary Cilia in Human Induced Pluripotent Stem Cell-Derived Neurons. Neurochem Res 44, 1736–1744 (2019). https://doi.org/10.1007/s11064-019-02806-4

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  • DOI: https://doi.org/10.1007/s11064-019-02806-4

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