ZFIN Image: Zhang et al., 2022, Figure 2

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Figure 2

adcy3a^-/^- and adcy5^-/^- double mutants show high melanosome aggregation. (A–H) adcy3a^-/^-;adcy3b^-/^- (B) and adcy6a^-/^-;adcy6b^-/^- (C) double mutants as well as adcy6a-MO-injected adcy3a^-/^- mutants (D) display normal distribution of melanosomes as that of WT embryos (A) under dark conditions at 4dpf. adcy6a-MO injected adcy5^-/^- mutants (F) exhibit comparable pigmentation as adcy5^-/^- (E). adcy3a^-/^-;adcy5^-/^- double mutants (G) develop a higher degree of aggregation of melanosomes than adcy5^-/^-, while co-injection of adcy3a and adcy5 mRNA can significantly rescue the defects in melanosome aggregation in adcy3a^-/^-;adcy5^-/^- double mutants (H). Scale bar = 100 μm in (A–H). (I) Pigmentation coverage shows no difference among wild-type embryos, adcy3a^-/^-;adcy3b^-/^-, adcy6a^-/^-;adcy6b^-/^-, and adcy3a^-/^- mutants injected with adcy6a-MO, as well as between adcy5^-/^- and adcy5^-/^- mutants injected with adcy6a-MO. adcy3a^-/^-;adcy5^-/^- double mutants have significantly reduced pigmentation areas compared to adcy5^-/^-, which can be rescued by co-injection of adcy3a and adcy5 mRNA (Student’s t-test, *** p < 0.001, **** p < 0.0001, n.s., not significant, p > 0.05). (J) Statistical analyses show that the number of melanocytes in adcy5^-/^- single mutants, and adcy3a^-/^-;adcy3b^-/^-, adcy6a^-/^-;adcy6b^-/^- and adcy3a^-/^-;adcy5^-/^- double mutants is comparable with that of WT embryos (Student’s t-test, n.s., not significant, p > 0.05). (K–R) WISH analyses showed the expression levels of sox10, mitfa, tyr, and dct are normal in adcy5^-/-;adcy3a^-/^- compared to WT embryos.

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Acknowledgments

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