Sheng-Hao Ding,Ying-Hui Bao,Jian-Hong Shen,Guo-Yi Gao,Yao-Hua Pan,Qi-Zhong Luo,Ji-Yao Jiang.[J].中华创伤杂志英文版,2016,19(1):16-24
Improved neurite outgrowth on central nervous system myelin substrate by siRNA-mediated knockdown of Nogo receptor
  
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KeyWord: RNA interferenceNeurite outgrowthNogo receptorGene knockdown
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Author NameAffiliation
Sheng-Hao Ding Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
Ying-Hui Bao Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
Jian-Hong Shen Department of Neurosurgery, Affiliated Hospital of Nantong University 
Guo-Yi Gao Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
Yao-Hua Pan Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
Qi-Zhong Luo Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
Ji-Yao Jiang Department of Neurosurgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University 
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Abstract:
      Purpose: To investigate the in vitro effect of short interfering RNAs (siRNAs) against Nogo receptor (NgR) on neurite outgrowth under an inhibitory substrate of central nervous system (CNS) myelin. Methods: Three siRNA sequences against NgR were designed and transfected into cerebellar granule cells (CGCs) to screen for the most efficient sequence of NgR siRNA by using reverse transcription polymerase chain reaction (RT-PCR) and immunofluorescence staining. NgR siRNA sequence 1 was found the most efficient which was then transfected into the CGCs grown on CNS myelin substrate to observe its disinhibition for neurite outgrowth. Results: Compared with the scrambled control sequence of siRNA, the NgR siRNA sequence 1 significantly decreased NgR mRNA level at 24 h and 48 h (p<0.05), which was recovered by 96 h after transfection. NgR immunoreactivity was also markedly reduced at 24 and 48 h after the transfection of siRNA sequence 1 compared with that before transfection (p<0.05). The NgR immunoreactivity was recovered after 72 h post-transfection. Moreover, the neurite outgrowth on the myelin substrate was greatly improved within 72 h after the transfection with siRNA sequence 1 compared with the scrambled sequence-transfected group or non-transfected group (p<0.05). Conclusion: : siRNA-mediated knockdown of NgR expression contributes to neurite outgrowth in vitro.
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