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Fig. 8 Testosterone directly acts on GPRC6A in zebrafish embryos. (A) Wild-type embryos at 3 h post-fertilization were exposed to vehicle, estradiol or testosterone. Cardiac edema was assayed at 3 days post fertilization. (B-E) Zebrafish with mutations in aromatase or nuclear estrogen receptors were exposed to testosterone and assayed for morphological phenotypes. Each data point is the mean percentage of embryos in a single clutch exhibiting cardiac edema (20-80 embryos per clutch). Clutches in the same treatment group were assayed on different days. Horizontal lines are the mean of each group. ns, not significant (P>0.05); one-way ANOVA with Tukey's multiple comparisons test.