Directing Astroglia from the Cerebral Cortex into Subtype Specific Functional Neurons
Figure 11
Reprogramming of postnatal astroglia expanded as neurosphere cells into functional GABAergic neurons following forced expression of Dlx2.
(A) Postnatal cortical astroglia were first expanded as neurospheres and subsequently transduced with a retrovirus encoding Dlx2 and DsRed. Double immunocytochemistry for DsRed (red) and vGaT (green) reveals that astroglia-derived neurosphere cells reprogrammed by Dlx2 exhibit a dense labelling of vGaT-positive puncta outlining their soma and their dendrites, indicating that Dlx2-reprogrammed neurons acquire a GABAergic identity (30 DPI). (B) High magnification view of a single astroglia-derived neurosphere cell transduced with Dlx2 illustrating the massive expression and the distribution of vGaT-positive puncta. The inset shows a high magnification view of the boxed area. (C–D) The fluorescence micrographs depict a fate-mapped, GFP-positive neuron (C, asterisk) co-expressing DsRed at 14 DPI (D, asterisk), indicating forced expression of Dlx2. (E) Step-depolarisation of the neuron shown in (C) and (D) evokes a GABAergic autaptic response (black trace) that is blocked by bicuculline (10 µM, red trace). Scale bars: A: 20 µm; B: 7 µm; Inset in B: 2 µm.