| First Author | Wohl SG | Year | 2016 |
| Journal | Glia | Volume | 64 |
| Issue | 5 | Pages | 743-62 |
| PubMed ID | 26732729 | Mgi Jnum | J:231095 |
| Mgi Id | MGI:5766848 | Doi | 10.1002/glia.22958 |
| Citation | Wohl SG, et al. (2016) miR-124-9-9* potentiates Ascl1-induced reprogramming of cultured Muller glia. Glia 64(5):743-62 |
| abstractText | The Muller glia of fish provide a source for neuronal regeneration after injury, but they do not do so in mammals. We previously showed that lentiviral gene transfer of the transcription factor Achaete-scute homolog 1 (Ascl1/Mash1) in murine Muller glia cultures resulted in partial reprogramming of the cells to retinal progenitors. The microRNAs (miRNAs) miR-124-9-9* facilitate neuronal reprogramming of fibroblasts, but their role in glia reprogramming has not been reported. The aim of this study was to test whether (1) lentiviral gene transfer of miR-124-9-9* can reprogram Muller glia into retinal neurons and (2) miR-124-9-9* can improve Ascl1-induced reprogramming. Primary Muller glia cultures were generated from postnatal day (P) 11/12 mice, transduced with lentiviral particles, i.e., miR-124-9-9*-RFP, nonsense-RFP, Ascl1-GFP, or GFP-control. Gene expression and immunofluorescence analyses were performed within 3 weeks after infection. Overexpression of miR-124-9-9* induced the expression of the proneural factor Ascl1 and additional markers of neurons, including TUJ1 and MAP2. When Ascl1 and miR-124-9-9* were combined, 50 to 60% of Muller glia underwent neuronal reprogramming, whereas Ascl1 alone results in a 30 to 35% reprogramming rate. Analysis of the miR-124-9-9* treated glial cells showed a reduction in the level of Ctdsp1 and Ptbp1, indicating a critical role for the REST pathway in the repression of neuronal genes in Muller glia. Our data further suggest that miR-124-9-9* and the REST complex may play a role in regulating the reprogramming of Muller glia to progenitors that underlies retinal regeneration in zebrafish. GLIA 2016;64:743-762. |
