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. 2022 Nov 23;8(47):eabq7219. doi: 10.1126/sciadv.abq7219

Fig. 7. The addition of Atoh1 to the IPA paradigm facilitates transition from a progenitor state to a differentiated neuron.

Fig. 7.

(A) Schematic of transgenic construct to express IPA with Atoh1 in MG. (B) Regeneration paradigm for inducing IPA:Atoh1 expression in MG in the damaged retina. (C) Representative immunofluorescence images of regenerated neurons from IPA:Atoh1 mice demonstrating MG-derived neurons (GFP+) are HuC/D+ (red) and not Otx2+ (purple). (D) Integrated UMAP of FACS-sorted MG-derived cells after regeneration paradigm with either IPA:Atoh1 or IPA-only overexpression. Highlighted in either blue or red are the RGC-like cells from each dataset that were subsetted for further comparative analysis. (E) Scatterplot highlighting differentially expressed genes between the RGC-like cells of the IPA:Atoh1 (blue) and IPA-only (red) regeneration paradigms. (F) GO analysis revealed that neurodevelopmental terms containing many retinal progenitor genes were down-regulated in the IPA:Atoh1 dataset versus IPA only. (G) Integrated UMAP of IPA:Atoh1 data as described above with previously generated Ascl1:Atoh1 dataset (13). Blue or red highlighting denotes RGC-like cells from each dataset compared in further analysis. (H) Scatterplot highlighting differentially expressed genes between the RGC-like cells of the IPA:Atoh1 and Ascl1:Atoh1 regeneration paradigms. (I) Bar plot of GO terms relating to neurite outgrowth enriched in the IPA:Atoh1 data. Known neuron projection genes listed are up-regulated with IPA:Atoh1 versus Ascl1:Atoh1. Mouse schematic was made with Biorender.com.