Fig. 6
From: TRPA1 exacerbates selective retinal ganglion cell vulnerability under acute ocular hypertension
2D and 3D observation of TRPA-mediated selective RGC damage pattern. (A) Schematic illustration of brain stereotaxic injection of CTB-488 and CTB-555 for grouping of RGCs. (B) Immunostaining of retina flatmount showing distribution of Plgn RGC and Mlgn RGC together with TRPA1 expression under control or AOH conditions. Scale bar: 50 μm. Plgn RGC and Mlgn RGC were retrogradely labeled by CTB. Note that in all other conditions where RGCs and their central projections were labeled with CTB, an intravitreal dose of CTB was applied for anterograde labeling. (C-D) Comparison of the relative TRPA1 intensity between Plgn RGC and Mlgn RGC in (C) control retina and (D) AOH retina (n = 10 for each group, *P < 0.05, N.S., non-significant). (E) 3D-reconstructed whole brain showing the changes of RGC central projection under AOH with or without conditional knock-out of Trpa1 in RGCs. Different brain regions were labeled, and the original data (left column) were surfaced (right column). AOH refers to the side of the brain where the RGCs of AOH model eye projected to. Scale bar: 1000 μm. (F) Heatmaps showing the relative value of the surface area, volume and sum of intensity of whole RGC central projection, OT, LGN, SC, and PrGPC, PrGMC in the AOH side compared with the control side in mice with or without conditional knock-out of Trpa1 in RGCs (n = 3 for each group, P values stand for differences between Trpa1fl/fl;Thy1-Cre− and Trpa1fl/fl;Thy1-Cre+ AOH side. *P < 0.05, **P < 0.02, ***P < 0.01). (G-H) Comparison of the extent of decrease in both the area, volume and sum of intensity between PrGPC and PrGMC (n = 3 for each experiment, paired t-test, **P < 0.02) in (G)Trpa1fl/fl;Thy1-Cre− and (H)Trpa1fl/fl;Thy1-Cre+ mice. (I) 3D-reconstructed whole brain. AOH refers to the side of the brain where the RGCs of AOH model eye projected to. Scale bar: 1000 μm. (J) Heatmaps showing the relative value of the surface area, volume and sum of intensity of different brain regions in the AOH side of brains compared with the control side with or without HC intravitreal injection (n = 3, *P < 0.05, ***P < 0.01 for comparison of AOH and AOH + HC group). Comparison of the extent of decrease in both the area, volume and sum of intensity between PrGPC and PrGMC (n = 3 for each experiment, paired t-test, *P < 0.05) in AOH mice without or with HC treatment were also shown (n = 3, *P < 0.05). (K) Direct comparison of PrGPC and PrGMC under AOH after HC treatment (n = 3 for each experiment, unpaired t-test, n.s., non-significant). Ar, surface area; CTB, cholera toxin subunit B; DCM, dichloromethane; LGN, lateral geniculate nucleus; OC, optic chiasm; ON, optic nerve; OT, optic tract; PrGMC, magnocellualr part; PrGPC, parvocellular part; SC, superior colliculi; SOI, sum of intensity; Vol, volume