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Fig. 7 GPRC6A antagonists rescue the testosterone phenotype in wild-type embryos but have no effect in gprc6a mutants. (A) Wild-type embryos were exposed to vehicle (0.1% DMSO) or GPRC6A antagonists: calindol (5 µM), NPS-2143 (5 µM) and epigallocatechin gallate (EGCG, 100 µM) with or without testosterone (30 µM), beginning at 3 h post-fertilization. (B) Homozygous zygotic gprc6a mutant embryos were exposed to vehicle, NPS-2143 or EGCG with or without testosterone. Morphological phenotypes were assayed at 3 days post-fertilization. Antagonists failed to reduce cardiac edema in gprc6a mutant embryos. Each data point is the mean percentage of embryos in a single clutch exhibiting cardiac edema (15-25 embryos per clutch). Clutches in the same treatment group were assayed on different days. Horizontal lines are the mean of each group. ****P<0.0001; ns, not significant (P>0.05); one-way ANOVA with Tukey's multiple comparisons test.