title: Lentivirus-Mediated RNA Interference Targeting RhoA Slacks the Migration, Proliferation, and Myelin Formation of Schwann Cells.
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.
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Jinkun
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Changhui
authors.initials: C
authors.lastname: Qian
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Mengjie
authors.initials: M
authors.lastname: Pan
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Xianghai
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Yuanyuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.|Center of Eletromicroscopy, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Yanmeng
authors.initials: Y
authors.lastname: Lu
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.|Center of Eletromicroscopy, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Zhitao
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Experimental Center, Shenyang Normal University, Shenyang, 110034, China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Yan
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Lixia
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China.
authors.forename: Zhongying
authors.initials: Z
authors.lastname: Liu
authors.affiliation: Department of Anatomy, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Pokfulam, Hong Kong, SAR, China.|State Key Laboratory of Brain and Cognitive Sciences, The University of Hong Kong, Pokfulam, Hong Kong, SAR, China.|Joint Laboratory of Jinan University and the University of Hong Kong, GHM Institute of CNS Regeneration, Jinan University, Guangzhou, 510632, China.
authors.forename: Wutian
authors.initials: W
authors.lastname: Wu
authors.affiliation: Department of Histology and Embryology, Southern Medical University, Guangzhou, 510515, China. jiasongguo@aliyun.com.|Key Laboratory of Tissue Construction and Detection of Guangdong Province, Guangzhou, 510515, China. jiasongguo@aliyun.com.|Institute of Bone Biology, Academy of Orthopedics, Guangdong Province, Guangzhou, 510665, China. jiasongguo@aliyun.com.
authors.forename: Jiasong
authors.initials: J
authors.lastname: Guo
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002465:Cell Movement / Q000502:physiology*; D049109:Cell Proliferation / Q000502:physiology*; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques / Q000379:methods; D018390:Gene Targeting / Q000379:methods; D016086:Lentivirus / Q000235:genetics; D009186:Myelin Sheath / Q000502:physiology*; D034622:RNA Interference / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012583:Schwann Cells / Q000502:physiology*; D020741:rho GTP-Binding Proteins / Q000172:deficiency* / Q000235:genetics
chemical_list: C000621224:RhoA protein, rat; D020741:rho GTP-Binding Proteins
keywords: Migration; Myelination; Peripheral nerve; Proliferation; RhoA; Schwann cell
references.citation: Antioxid Redox Signal. 2014 May 1;20(13):2026-42
references.pmid: 24251358
references.citation: Endocrinology. 2011 May;152(5):2037-47
references.pmid: 21385940
references.citation: Dev Biol. 1973 Jun;32(2):401-16
references.pmid: 4789698
references.citation: Neuroscientist. 2016 Jun;22(3):252-65
references.pmid: 25686621
references.citation: Eur J Neurosci. 2004 Jun;19(12 ):3142-50
references.pmid: 15217370
references.citation: Am J Physiol Heart Circ Physiol. 2013 Feb 1;304(3):H369-81
references.pmid: 23203966
references.citation: Biochim Biophys Acta. 2014 Nov;1842(11):2266-75
references.pmid: 25108283
references.citation: Nat Rev Neurosci. 2005 Sep;6(9):683-90
references.pmid: 16136172
references.citation: Cold Spring Harb Perspect Biol. 2015 Jun 08;7(8):a020529
references.pmid: 26054742
references.citation: J Neurosci. 2004 Apr 21;24(16):3953-63
references.pmid: 15102911
references.citation: Glia. 2013 Nov;61(11):1906-21
references.pmid: 24014231
references.citation: Mol Cell Neurosci. 2007 May;35(1):120-9
references.pmid: 17382558
references.citation: J Neuropathol Exp Neurol. 2003 May;62(5):520-9
references.pmid: 12769191
references.citation: Drug Resist Updat. 2015 Mar;19:22-32
references.pmid: 25660168
references.citation: Mol Biol Evol. 2007 Jan;24(1):203-16
references.pmid: 17035353
references.citation: Exp Neurol. 2010 Nov;226(1):47-57
references.pmid: 20696161
references.citation: J Cell Biochem. 2011 Dec;112(12):3786-96
references.pmid: 21826705
references.citation: Glia. 2015 Aug;63(8):1376-93
references.pmid: 25921593
references.citation: Arch Oral Biol. 2014 Feb;59(2):149-57
references.pmid: 24370186
references.citation: Alcohol. 2013 Aug;47(5):417-26
references.pmid: 23731693
references.citation: Ann Neurol. 2014 Feb;75(2):255-65
references.pmid: 24339129
references.citation: Curr Mol Med. 2014 Feb;14(2):199-208
references.pmid: 24467208
references.citation: Cell Signal. 2014 Dec;26(12):2998-3006
references.pmid: 25286227
references.citation: Glia. 2013 Dec;61(12 ):2009-22
references.pmid: 24115248
references.citation: Front Cell Neurosci. 2013 Sep 24;7:161
references.pmid: 24065889
references.citation: J Neurosci. 2012 Nov 28;32(48):17251-61
references.pmid: 23197717
references.citation: J Cell Biol. 2007 Jun 18;177(6):1063-75
references.pmid: 17576799
references.citation: J Neurosci. 2010 Mar 17;30(11):4120-31
references.pmid: 20237282
references.citation: J Cell Biol. 2007 Jun 18;177(6):1051-61
references.pmid: 17576798
references.citation: Nat Protoc. 2014 May;9(5):1160-9
references.pmid: 24762783
references.citation: Nanomedicine. 2013 Apr;9(3):305-15
references.pmid: 22960189
references.citation: Handb Clin Neurol. 2013;115:55-79
references.pmid: 23931775
references.citation: Cold Spring Harb Perspect Biol. 2015 Aug 03;7(8):a005959
references.pmid: 26238352
references.citation: Mol Med Rep. 2014 Jan;9(1):173-9
references.pmid: 24172987
references.citation: Exp Neurol. 2003 Sep;183(1):34-46
references.pmid: 12957486
references.citation: Brain Res. 1979 Apr 6;165(1):105-18
references.pmid: 371755
references.citation: Proc Natl Acad Sci U S A. 2004 Jun 8;101(23):8774-9
references.pmid: 15161978
references.citation: Mol Cell Neurosci. 2005 Mar;28(3):509-23
references.pmid: 15737741
references.citation: J Cell Biol. 2007 Jun 18;177(6):953-5
references.pmid: 17576794
references.citation: J Cell Sci. 2014 Jul 15;127(Pt 14):2999-3004
references.pmid: 25024457
references.citation: Neural Regen Res. 2014 Dec 15;9(24):2132-41
references.pmid: 25657734