Mechanism of AAV9-OPTN on autophagy pathway in atrophic lateral sclerosis model mice
Objective To explore the effect of self complementary double stranded adenovirus 9 mediated optineurin(AAV9-OPTN)on the autophagy pathway in a transgenic mouse model of amyotrophic lateral sclerosis(ALS)SOD1-G93A.Methods The animal model of ALS,transgenic SOD1-G93A mutant mice were used.At 30-40 days after birth,they were randomly assigned to the treatment group and control group.The treatment group received intrathecal injection of AAV9-PTN,while the control group received intrathecal injection of AAV9-GFP.After 40-50 days of Oinjection,immunoblotting technology was used to detect changes in the content of OPTN in the lumbar spinal cord.Immunofluorescence staining was used to observe the localization of OPTN,while immunofluorescence staining was used to observe the staining and fluorescence content changes of autophagy pathway related proteins p62,TBK1,LC3.Results The immunoblotting showed that after intrathecal injection of AAV9-OPTN,the protein content of OPTN in the lumbar spinal cord was significantly higher than that in the GFP control group,and the fluorescence intensity of immunofluorescence staining for OPTN was significantly higher than that in the solvent control group(allP＜0.01).The immunofluorescence staining results of autophagy pathway proteins showed that the fluorescence intensity of TBK1 and LC3 in the AAV9-OPTN treatment group were significantly higher than that in the control group,while the fluorescence intensity of p62 decreased,showing statistical differences compared to the control group.Conclusion AAV9-OPTN can be transferred into the lumbar spinal cord of mice and induced the expression of OPTN protein,which can activate the autophagy pathway in the SOD1-G93A transgenic mouse model.
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