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.
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
You-Ming
YM
Fan
Chi-Pui
CP
Pang
Alan R
AR
Harvey
Qi
Q
Cui
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