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： Th...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 ceils （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 I was found the most efficient which was then transfected into the CGCs grown on CNS myelin substrate to observe its disinhihition for neurite outgrowth. Results： Compared with the scrambled control sequence of siRNA, the NgR siRNA sequence 1 signifi- cantly decreased NgR mRNA level at 24 b 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 tile transfection of siRNA sequence 1 compared with that before transfection （p 〈 0.05）. The NgR immunoreactiviW was recovered after 72 h post-transfection. Moreover, the neurite outgrowth on tire 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.展开更多
文摘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 ceils （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 I was found the most efficient which was then transfected into the CGCs grown on CNS myelin substrate to observe its disinhihition for neurite outgrowth. Results： Compared with the scrambled control sequence of siRNA, the NgR siRNA sequence 1 signifi- cantly decreased NgR mRNA level at 24 b 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 tile transfection of siRNA sequence 1 compared with that before transfection （p 〈 0.05）. The NgR immunoreactiviW was recovered after 72 h post-transfection. Moreover, the neurite outgrowth on tire 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.
