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Fig. 6

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ZDB-IMAGE-200422-7
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Figures for Mackin et al., 2019
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Figure Caption

Fig. 6 Plasticity of lws differential expression by TH (T4) in normal and athyroid juveniles. (A) Schematic of experimental workflow including thyroid ablation at 2 dpf followed by T4 treatment starting at 26 dpf for 5 d, of half the athyroid group and half of the controls. (B and C) qPCR of lws1 (A) and lws2 (B) transcript abundance. Scatter plots indicate fold-change abundance (2−ddCT) of the indicated transcripts. Colors of dots correspond to separate experiments. Each dot represents a biological replicate (2 retinas from an individual fish). lws1 abundance (B) in DMSO (control) (n = 9), is unchanged by T4 GOF (n = 9) P = 0.21 compared to control, decreased in athyroid (LOF) (n = 6) P = 0.006 compared to control, and increased in Rescue (LOF + T4) (n = 7) P = 0.006 compared to LOF. lws2 abundance (C) in control (n = 9), is decreased by T4 GOF (n = 9) P = 2.66e-06 compared to control, increased in LOF (n = 6) P = 4.38e-02 compared to control, and decreased in Rescue (n = 7) P = 3.61e-08 compared to LOF. ddCT values were calculated by subtracting the dCT value for each fish from the average dCT values from the control fish in each experiment. P values were calculated by comparing the ddCT values for each fish from each condition using the Kruskal–Wallis test and the Conover post hoc test further adjusted by the Benjamini–Hochberg false-discovery rate method. Statistical notation: *P < 0.05, **P < 0.01, ****P < 0.0001. (D–G) lws:PAC(H) whole-mounted retinas visualized by confocal microscopy of 31 dpf DMSO (control) (D), T4 (GOF) (E), athyroid (LOF) (F), and Rescue (LOF+T4) (G) juveniles. D, dorsal; N, nasal. Arrowhead in F indicates restricted ventral expression domain of lws1 in athyroid juvenile retina. (H) Whole-mounted retina of an age-matched RARE:eYFP juvenile showing domain of RA signaling restricted to ventral retina (arrowhead), and matching the domain of GFP (lws1) expression in athyroid retinas (arrowhead in F) of juvenile fish. (I) High-magnification view of lws:PAC(H) retina showing GFP/RFP colabeling (opsin switching) in T4-treated juveniles (not the same retina pictured in E). Green arrowhead indicates a GFP+ cone. White arrowhead indicates a colabeled (GFP+/RFP+) cone. (Scale bar in D [applies to D–H], 100 μm; in I, 50 μm.)

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