Figure 5

Retinal immunohistochemistry and OCT image of adult zebrafish

Panels (A) and (B) present the expression and localization of the Panx1a (in green) antibody throughout the retinal layer. The Panx1a is localized in the horizontal cell layer (HC) and in the ON/OFF ganglion cell layer, as it is shown in panel (a1). The Panx1a-KO served as the negative control (scale bars, 25 μm). Panel (C) is a B-mode OCT image of the TL zebrafish retina obtained using our modified 880 nm OCT system and zebrafish retina probe, and Panel (D) is a B-mode retina OCT of Panx1a−/−. The red square in Panel (C) indicates the region selected by the custom MATLAB code for the analysis and measurement of each layer's thickness. Panel (E) shows the fitted plot of the retinal layers. The green line shows the thickness of the GCL layer, and the yellow line presents the whole retina thickness. Panels (F and G) present estimation plots of whole-retina thickness and ganglion cell layer (GCL) thickness in the 8-month-old group, demonstrating a significant decrease in both measurements (n = 30 for Panx1a^+/+ and KO, Statistical analysis: Welch’s t test; p < 0.05). Panel (H) is showcasing an en-face OCT image of Panx1a^+/+ retina. Panel (I) is an en-face OCT image of Panx1a^−/− retinal vessels, with the red rectangle highlighting the "tortuous vessel" phenotype observed in a few Panx1a^−/− eyes. Statistical analysis: Welch’s t test; Significance ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, and ∗∗∗∗p < 0.0001. Scale bars, 500 μm.