Expression Of Miz1 In Alzheimer's Disease And Its Relationship With Autophagy

Objective:Our previous RNA-sequencing analysis in acutely isolated neurons has revealed that transcription factor Miz1 was differential expressed in APP/PS1 mice,compared with age-matched control littermates.In this study,the relationship between Miz1 and AD was studied by Western blotting,qPCR and immunofluorescence.Methods and Results:1.Elevated Miz1 expression in the cortex of APP/PS1 mice,compared with age-matched WT mice.Western blot analysis was conducted to determine the expression levels of Miz1,the expression level of Miz1 was higher in the cortex of AD mice than in that of the WT mice.Meanwhile,There was no significant difference between the two types of mice in hippocampal staining.The same results were detected by qPCR and Immunohistochemistry.2.To explore the relationship between Miz1 neurons and astrocytes in the brains of WT and AD mice,we observed the location of Miz1 in the WT and AD mouse brain by immunofluorescence assay.We performed double-label immunofluorescence stainings with antibodies against Miz1,with the neuronal marker MAP2 and with the astrocyte marker GFAP.The images showed that Miz1(green)was co-expressed with GFAP(red)in both the cortex and hippocampus of WT and AD mice.We found that Miz1(green)was co-expressed with MAP2(red)in the brains of WT and AD mice.3.The plasmid PCMV-Miz1 was transiently transfected into HT22 cells.After 24 hours of treatment,the cells were extracted for Western blotting.And the expression level of APP metabolism related protein was decreased and the expression of autophagy related protein was increased in the Miz1 treated group compared with the same group transfected with empty plasmid.These results suggested that Miz1 may play a role in inhibiting APP metabolism and promoting autophagy at the protein levelConclusions:Elevated Miz1 expression in the cortex of AD mice compared with age-matched WT mice.Miz1 was co-expressed with neurons and astrocytes in the brains of WT and AD mice.Mzi1 may be involved in the regulation of APP metabolism and autophagy.