Journal: Neural Regeneration Research

Article Title: Selective deletion of zinc transporter 3 in amacrine cells promotes retinal ganglion cell survival and optic nerve regeneration after injury

doi: 10.4103/1673-5374.373660

Figure Lengend Snippet: Specific knockout of ZnT3 in ACs and RGCs. (A) Schematic illustration of the ZnT3 ( Slc30a3 ) knockout strategy by CRISPR/Cas9 technology. (B) Schematic of the retinal structure (left); VGLUT2-Cre mice selectively express Cre in RGCs (green), whereas VGAT-Cre mice selectively express Cre in amacrine (red) and horizontal cells (black). Synaptic connections between RGCs and ACs are shown on the right; Zn 2+ is encapsulated in the synaptic vesicles in ACs, ZnT3 transports the vesicles to the synaptic cleft, and Zn 2+ released into RGCs. (C) Genotyping of VGAT-Cre (mut/wt), Slc30a3 fl/fl , and Slc30a3 fl/fl VGAT-Cre mice ( n = 4). The white boxes indicate the genotyping result of Slc30a3 fl/fl VGAT-Cre mice. VGAT-Cre: Mutant = 200 bp, wt = 323 bp; Slc30a3 : fl = 304 bp, wt = 202 bp. (D) Genotyping of VGLUT2-Cre (mut/wt), Slc30a3 fl/fl and Slc30a3 fl/fl VGLUT2-Cre mice ( n = 4). The white boxes indicate the genotyping result of Slc30a3 fl/fl VGLUT2-Cre mice. VGUT2-Cre: Mutant = 124 bp, wt = 245 bp; Slc30a3: fl = 304 bp, wt = 202 bp. AC: Amacrine cell; CRISPR/Cas9: clustered regularly interspaced short palindromic repeats/CRISPR-associated protein 9; RGCs: retinal ganglion cells; wt or wt/wt: C57BL/6J wild-type mice; VGAT: vesicular γ-aminobutyric acid transporter; VGLUT2: vesicular-glutamate transporter 2; ZnT3: zinc transporter 3.

Article Snippet: Genotyping was performed by polymerase chain reaction using DNA polymerase (Vazyme, Nanjing, China, Cat# P222).

Techniques: Knock-Out, CRISPR, Mutagenesis