Marked effect of RhoA-specific shRNA-producing plasmids on neurite growth in PC12 cells.
In order to promote neurite outgrowth, we constructed a plasmid producing RhoA-specific small hairpin RNA (shRNA) to block RhoA expression and tested its actions in PC12 cells. Our results show that the shRNA-mediated RNA interference (RNAi) successfully suppressed RhoA expression. As a consequence of RhoA knockdown, F-actin levels were significantly reduced and processes were markedly induced. These processes express two neurite markers, neurofilament and betaIII tubulin. This study demonstrates that plasmid-producing shRNA specific for RhoA can act as an effective tool to induce neurite outgrowth and further confirms the neurite growth-inhibitory role of RhoA. This shRNA may thus be a useful tool in promoting neurite outgrowth and could be applicable in neural repair after CNS injury.
Fan
You-Ming
YM
Joint Shantou International Eye Center of Shantou University and The Chinese University of Hong Kong, Shantou University Medical College, Shantou, PR China. youmingf@yahoo.com.cn
Pang
Chi-Pui
CP
Harvey
Alan R
AR
Cui
Qi
Q
D000199:Actins / Q000096:biosynthesis / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D016501:Neurites / Q000502:physiology*; D016900:Neurofilament Proteins / Q000096:biosynthesis / Q000378:metabolism; D016716:PC12 Cells; D010957:Plasmids / Q000235:genetics*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000502:physiology; D014404:Tubulin / Q000096:biosynthesis / Q000378:metabolism; D020742:rhoA GTP-Binding Protein / Q000235:genetics* / Q000378:metabolism / Q000502:physiology
D000199:Actins; D016900:Neurofilament Proteins; D034741:RNA, Small Interfering; C488416:Tubb3 protein, rat; D014404:Tubulin; D020742:rhoA GTP-Binding Protein