Improved neurite outgrowth on central nervous system myelin substrate by siRNA-mediated knockdown of Nogo receptor

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• 作者：Sheng-Hao Ding^a ; Ying-Hui Bao^a ; ^{bombbao@126.com" class="auth_mail" title="E-mail the corresponding author} ; Jian-Hong Shen^b ; Guo-Yi Gao^a ; Yao-Hua Pan^a ; Qi-Zhong Luo^a ; Ji-Yao Jiang^a ; ^{jiyaojiang@126.com" class="auth_mail" title="E-mail the corresponding author}
• 关键词：RNA interference ; Neurite outgrowth ; Nogo receptor ; Gene knockdown
• 刊名：Chinese Journal of Traumatology (English Edition)
• 出版年：2016
• 出版时间：February 2016
• 年：2016
• 卷：19
• 期：1
• 页码：16-24
• 全文大小：2093 K

文摘

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.