Effect of vimentin as a glycosylation molecule on neural differentiation
Objective To explore the role of vimentin as a glycosylation molecule in neural differentiation of PC12 cells.Methods RNA interference plasmid to silence vimentin expression and recombinant plasmid with high expression of vimentin were constructed.The expression of vimentin was detected by Western blot.PC12 cells were transfected with plasmid empty vector,high-expression plasmid or interference plasmid respectively and cultured for 3 days with or without Cyclo-ManN propanyl perac(Cyclo-ManN pro)pretreatment.The morphology of cells was observed and recorded.The neurite length and percentage of differentiated cells were measured for quantitative differentiation analysis.The interaction between vimentin and biotinylated Maackia amurensis lectin Ⅱ(MAL Ⅱ)was immunoprecipitated in PC12 cells with or without Cyclo-ManN pro pretreatment.The glycosylation of vimentin was vertified.Results The interfering and high expressed vimentin recombinant plasmids were constructed and identified successfully.Compared with plasmid empty vector,high-expression plasmid increased the neurite length and percentage of differentiated cells,while interference plasmid decreased the above indicators in PC12 cells with or without Cyclo-ManN pro pretreatment(P＜0.05 or P＜0.01).Compared with PC12 cells without Cyclo-ManN pro pretreatment,PC12 cells pretreated by Cyclo-ManN pro had significantly more vimentin recognized by MAL Ⅱ.Conclusion Vimentin may act as glycosylation molecule to promote neural differentiation of PC12 cells.
NETL
NSTL
万方数据
