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Lentivirus-Mediated RNA Interference Targeting RhoA Slacks the Migration, Proliferation, and Myelin Formation of Schwann Cells

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Abstract

RhoA, a member of Rho GTPases family, is known to play an important role in remodeling actin cytoskeleton. During the development of the peripheral nervous system (PNS), Schwann cells undergo proliferation, migration, and radial sorting and finally wrap the related axons compactly to form myelin sheath. All these processes involve actin cytoskeletal remodeling. However, the role of RhoA on Schwann cell during development is still unclear. To address this question, we first used a lentiviral vector-mediated short hairpin (sh) RNA targeting RhoA to knock down the expression of RhoA in the cultured Schwann cells in vitro. Effects of RhoA on Schwann cell proliferation and migration were examined by BrdU assay and transwell assay, respectively. Results of the present study indicated that downregulated RhoA expression in cultured Schwann cells significantly slacked the cells’ capabilities of migration and proliferation. Then, we investigated the role of RhoA in the developing rat sciatic nerves. Immunohistology and Western blotting showed that RhoA was mainly expressed in Schwann cells in the sciatic nerves and was peaked at 2 weeks postnatal then kept in low level up to 8 weeks. In the subjected rats whose sciatic nerves were microinjected with lentiviral vectors at postnatal 3 days, we found that the lentiviruses mainly transfected Schwann cells, and the RhoA expression in the transfected Schwann cells was significantly knocked down. Four weeks after lentivirus microinjection, immunohistology and transmission electron microscopy illustrated that RhoA knockdown resulted in hypomyelination and significant decrease of the thickness of myelin in the transfected area. Overall data of current study suggested that RhoA plays a critical role in Schwann cell biology and is essential for myelination in developing peripheral nerve.

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Acknowledgments

This work is supported by the National Key Basic Research Program of China (2014CB542202 and 2014CB542205), National Natural Science Foundation of China (81371354 & 81571182), Science and Technology Foundation of Guangdong Province (2015A020212024), and Natural Science Foundation of Guangdong Province (S2013010014697) to J Guo and Hong Kong SCI Fund to W Wu.

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Authors and Affiliations

  1. Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China

    Jinkun Wen, Changhui Qian, Mengjie Pan, Xianghai Wang, Yuanyuan Li, Yanmeng Lu, Zhitao Zhou, Lixia Li, Zhongying Liu & Jiasong Guo

  2. Center of Eletromicroscopy, Southern Medical University, Guangzhou, 510515, China

    Yanmeng Lu & Zhitao Zhou

  3. Experimental Center, Shenyang Normal University, Shenyang, 110034, China

    Qing Yan

  4. Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, SAR, China

    Wutian Wu

  5. State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong, Pokfulam, Hong Kong, SAR, China

    Wutian Wu

  6. Joint Laboratory of Jinan University and the University of Hong Kong, GHM Institute of CNS Regeneration, Jinan University, Guangzhou, 510632, China

    Wutian Wu

  7. Key Laboratory of Tissue Construction and Detection of Guangdong Province, Guangzhou, 510515, China

    Jiasong Guo

  8. Institute of Bone Biology, Academy of Orthopedics, Guangdong Province, Guangzhou, 510665, China

    Jiasong Guo

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  1. Jinkun Wen
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Correspondence to Jiasong Guo.

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Wen, J., Qian, C., Pan, M. et al. Lentivirus-Mediated RNA Interference Targeting RhoA Slacks the Migration, Proliferation, and Myelin Formation of Schwann Cells. Mol Neurobiol 54, 1229–1239 (2017). https://doi.org/10.1007/s12035-016-9733-5

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