Fig 3

RGC axon volume and trajectory examination in gsx1 mutants.

(A) Example max projection of a confocal z-stack in wildtype and (B)gsx1^y689, with Tg(atoh7:eGFP) anti-GFP labeling. Dashes around axons indicates regions of interest and mask application. (Aⁱ-Bⁱ) Example 3D volume rendering taken from Imaris when Labkit is used. (C) Box and whisker plot with individual data points included for volumes. There was a significant difference in volume of RGC axons between wildtype (M = 904100.00μm³, SD = 134984.32, SEM = 42685.79μm³, n = 10) and gsx1 mutants (M = 564000.00μm³, SD = 80067.47, SEM = 24141.25μm³, n = 11); t(19) = 7.1, p < 0.001. Post-hoc power analysis = 100%. Colored dots correspond to Fig 2 images (magenta), and D-Eⁱⁱ representative minimum (yellow), maximum (light gray), and average (cyan) axon volume images. (D-Dⁱⁱ) Wildtype and (E-Eⁱⁱ) mutant RGC axon images that have been depth color coded to show AF9 in blue as the deepest AF. These images represent the RGC axon phenotype variability in wildtypes and mutants as indicated in panel C. (F-Gⁱⁱⁱ) Max projection of confocal z-stacks in wildtypes (F-Fⁱⁱⁱ) and gsx1^y689(G-Gⁱⁱⁱ), with DiI (magenta) injected into the temporal retina and DiO (green) injected into the nasal retina. Scalebar = 100μm. (Fⁱⁱⁱand Gⁱⁱⁱ) Zoomed in pretectal region with consistent loss of AF7 in gsx1^y689. Scalebar = 50μm.