Figure 1
From: Heterogeneous brain region-specific responses to astrocytic mitochondrial DNA damage in mice
Mito-PstI depletes and deletes astrocytic mtDNA in vivo. (A) Schematic showing the AAV 2/5 vectors for stereotaxic injection into specific brain regions. GfaABC1D: Gfap promoter. Mito-7: COX8A mitochondrial signal sequence. 3XGGS: 12 amino acid linker. (B) Images depicting Mito-GFP fluorescence on NeuN stained HPC and DLS tissue, respectively. GFP expression was highly astrocyte specific. Scale bar = 500 μm. (C) Diagram for mtDNA genome with location of PstI sites (red bars), PCR primers for Cytochrome b (white arrows), and for detection of possible deletions (black and gray arrows). Nucleotide (nt) positions are indicated. (D) Relative mtDNA content quantification showing a ~ 40% reduction in mtDNA content in Mito-PstI-treated tissue. Each data point is midbrain homogenate from a single, male mouse. Errors bars are ± SEM, p-value for the relative mtDNA content quantification is based on a two-sample t-test, n = 3 male mice per condition. (E) Detection of potential deletions. Size markers in the right-most lane; ~ 700 bp fragment, suggestive of mtDNA recombination, only observed in Mito-PstI treated samples (n = 3/condition).
