Objective To investigate the effect ofspecific knockdown of the Wnt3a gene at embryonic day 13(E13)on brain neurogenesis in mice.Methods RT-qPCR was used to verify the knockdown rate of Wnt3a in neural stem cells in the cerebral cortex,and intrauterine electroporation was used to transfect Wnt3a-shRNA plasmid into the cerebral cortex of fetal mice at E13 to downregulate the expression of Wnt3a in neural stem cells in the cerebral cortex.The mouse brain tissue samples were collected at embryonic day 16(E16),and immunofluorescent staining was performed to observe the changes in neuronal development after the downregulation of Wnt3a.Results RT-qPCR showed that the mRNA expression level of Wnt3a in neural stem cells was reduced by 54％after knockdown,with a significant difference compared with the control group(t=11.260,P＜0.001).Immunofluores-cent staining showed that the downregulation of Wnt3a led to a reductionin the proportion of green fluorescent protein(GFP)-posi-tive cells in the cortical plateof the cerebral cortexand an increase in the proportion of GFP-positive cells in the intermediate zone(IZ)(t=4.400,4.482;P＜0.01).Conclusion The specific downregulation of the Wnt3a gene in the brain of mice at E13 can lead to an aberrant distribution of GFP fluorescence,thereby disrupting the process of neurogenesis.
维普
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