1. RPE dedifferentiation and mesenchymal transition after PGC-1 deletion.
RPE dedifferentiation and mesenchymal transition after PGC-1 deletion. (A) Immunodetection of mitochondrial and EMT markers on cryosections of eyes collected 4 mo postinjection with AAV-GFP or AAV-Cre, showing marked loss of the ECT component COXIV in the RPE layer (defined by the white dotted lines) of the AAV-Cre–injected animals, whereas the pro-EMT transcription factors TWIST1 and ZEB1 were strongly up-regulated. Mesenchymal phenotypic transition is further demonstrated by increased collagen VI deposition and vimentin perinuclear condensation. Scale bars are 20 μm. (B) RPE flat-mount preparation immunostained for F-actin or ZO-1 (red), demonstrating increased multinucleated cells (arrowheads) with large pigmented vacuoles (arrows) and loss of membranous ZO-1 distribution. Nuclei were costained with DAPI (Blue). Scale bar is 50 μm. (C) Percentage of mononucleated, binucleated, and polynucleated (>2 nuclei/cell) cells quantified on F-actin stained RPE flat mounts (n = 6). Error bars are means ± SEM. Data were analyzed by unpaired t test. **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ compared with AAV-GFP group.
Mariana Aparecida Brunini Rosales et al. LSA 2019;2:e
© 2019 Rosales et al.