Fig. 1

Only oncogenic GNAQ^Q209P is sufficient to induce choroidal melanocyte hyperplasia. A. Schematic representation of elements in the Tol2-based transposon vector driving the expression of oncogenic GNAQ^Q209P under the control of zebrafish mitfa promoter in the melanocyte lineage and Venus fluorescent reporter under the control of cryaa promoter in the eye lens. Abbreviations: ITR, inverted terminal repeat. B. Example of a 5 dpf transgenic zebrafish embryo with a fluorescent eye lens. Scale bar, 100 μm. C. RT-qPCR data showing a 4.1 fold increase in GNAQ expression in the melanocytes of 2-month-old F₁ Tg (mitfa:GNAQ^Q209P) zebrafish, as compared to non-injected controls. Data represents mean ± SEM of triplicates of three independent experiments. *** P <0.05 using two-tailed, unpaired t test. D, E, F, G. H&E staining of transverse sections of formalin-fixed and paraffin-embedded eye specimens of control wild-type, Tg (mitfa:GNAQ^Q209P), Tg (mitfa:NRAS^Q61L), and Tg (mitfa:BRAF^V600E) zebrafish, respectively. Choroidal hyperplasia observed in the thickened choroid (E; black arrowhead) was only observed in transgenic animals expressing oncogenic GNAQ^Q209P. In contrast, as compared to control wild-type H. and Tg (mitfa:GNAQ^Q209P) I. hyperplasia of cutaneous melanocytes (black arrowhead) was only detected in transverse sections of the torso region of Tg (mitfa:NRAS^Q61L) J. and Tg (mitfa:BRAF^V600E) K. zebrafish. Abbreviations: RPE, retinal pigmented epithelium. Scale bars, 20 μm.