Figure 4.

Transcript levels of pineal gland markers are greatly reduced in adult bsx ^−/− mutant zebrafish. Analyses of transcript levels of bsx (A) aanat2 (B), agrp2 (C), exorh (D), otx5 (E), and crx (F) at daytime (ZT6) and nighttime (ZT18) as determined by qRT‐PCR. Copy numbers were normalized to the geometric means of copy numbers of eef1a1 and actb2, which were detected at high and stable levels in all samples; copy numbers were further divided by 100 to account for abundant transcript levels. Two‐way ANOVA detected significant effects of time of day on transcript levels of aanat2 (p < .0001) and significant effects of genotype on transcript levels of bsx (p < .0001), aanat2 (p < .0001), agrp2 (p < .0001), exorh (p < .0001), otx5 (p < .001), and crx (p < .001). Transcript levels of homozygous (bsx ^−/−) and heterozygous (bsx ^{+/ −}) mutants were compared to those of wildtype (bsx ^+/+) controls. Values on the graphs represent mean± SEM; n = 4, where each sample is pooled tissue from three animals. p‐values as determined by multiplicity‐adjusted Dunnett's multiple comparisons tests: ns, not significant, p > .05; *, p < .05; **, p < .01; ***, p < .001; ****, p < .0001. ZT, Zeitgeber time