In Vivo Reprogramming of Resident NG2 Glia in a DA-Denervated Striatum and in the Midbrain
(A and B) Confocal images (stitched into a tile) of striatal sections showing GFP+ reprogrammed neurons that occur with similar efficiency (A and B) and morphology (A′ and B′) in the intact (A) and 6-OHDA lesioned (B) brain.
(C and D) Neurons in both conditions, (C) intact and (D) lesioned, showed repetitive current-induced action potentials (AP; traces on the left) and spontaneous postsynaptic events (traces on the right), in the absence of any drugs or stimulation.
(E) Reprogrammed neurons in the intact brain express GFP (E′), but not TH (E″).
(F) TH+ neurons were observed in the striatum after 6-OHDA mfb lesion, but these cells do not co-express GFP (arrows).
(G) Confocal image of abundant TH+ cells appearing in control animals (lesioned but no reprogramming factors).
(H) Quantification of the average number of TH+ cells found per section in control lesioned versus lesioned + reprograming factors groups (n = 6/group).
(I–L) Analysis of TH+ cells show that they do not express the striatal markers GAD65/67 (I) or PV (JK), but do express Nurr1 (K), and low levels of DAT (L).
(M–M″) In vivo reprogramming of resident NG2 glia in the midbrain results in endogenous TH+ cells (red) intermingled with GFP+ (green) reprogrammed neurons, with no overlap.
Data are presented as means ± SEM. Scale bars: (A–F) 100 μm; (A′ and B′) 100 μm; (G) 50 μm; (I–L) 25 μm; (M–M″) 50 μm. See also Figure S3.