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. 2022 Sep 19;10:914386. doi: 10.3389/fcell.2022.914386

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Copyright © 2022 Le, Appel, Pannullo, Hoang and Blackshaw.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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GFAP promoter constructs expressing NeuroD1, Mybl1 and Insm1 show ectopic mCherry expression in amacrine and ganglion cells. Representative immunostaining for GFP, mCherry and 3 neuronal markers: (A) Tfap2a, (B) Rbpms, and (C) Otx2 expression in the retinas collected 21 days post GFAP AAV infection. GFAP-NeuroD1-mCherry, GFAP-Mybl1-mCherry and GFAP-Insm1-mCherry showed little to no colocalization of construct-derived mCherry and Müller glia-specific Sun1-GFP. High levels of mCherry expression in amacrine and ganglion cells, but not in bipolar cells were observed in retinas infected with these GFAP AAV constructs. Yellow arrowheads indicate co-labeled mCherry+ & marker + cells. Quantification of transduction efficiency (mCherry+ & GFP + cells/GFP + cells) (D) and transduction specificity (mCherry+ & GFP + cells/mCherry + cells) (E). Quantification of mean percentage ±SD of marker & mCherry+/GFP cells: Tfap2a+ & mCherry+/GFP + cells (in INL and GCL), Rbpms+ & mCherry+/GFP + cells (in GCL) and Otx2+&mCherry+/GFP + cells (in INL) (F). Significance was determined via one-way ANOVA or two-way ANOVA with Dunnett’s test: ****p < 0.0001. Each data point in the bar graphs was calculated from an individual retina (n = 6). INL, inner nuclear layer; GCL, ganglion cell layer. Scale bar = 40 μm.