Knockout of protein kinase C interacting protein 1 gene induces neuronal oxidative stress
Objective To investigate the effect of protein kinase C interacting protein 1(PICK1)knockout on neuronal oxida-tive stress response,and to clarify the role of PICK1 in neuronal oxidative stress response.Methods The mouse tail of 1cm was trimmed and homogenized,then DNA and proteins were extracted,and the genotypes of wild-type mice and PICK1 knockout mice were identified using PCR and western blot.The level of reactive oxygen species(ROS)was detected by ELISA and glutathione(GSH)assay kit was used to exam GSH level.Western blot was operated to check the expression level of oxidative damage indicator Nitrotyrosine,and immu-nofluorescence staining was done to detect the co-location of Nitrotyrosine and MAP2,the marker of neurons in the mouse brain.Results The wild-type mice,PICK1 gene heterozygous mice and homozygous mice were successfully identified.The levels of oxidative stress factors ROS(P＜0.05)and Nitrotyrosine in the brain of PICK1 knockout mice significantly increased(P＜0.05),while the levels of antioxidant stress factor GSH significantly decreased(P＜0.05).The immunofluorescence co-staining results of Nitrotyrosine and MAP2 showed oxidative damage to neuronal cells in PICK1 knockout mice.Conclusion Knockout of PICK1 gene could lead to oxidative stress response in mouse neurons and suggested that PICK1 may be a target for inhibiting neuronal oxidative stress.
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维普
