Fig. 3

rbpms2;dmrt1TMs are sterile, unlike rbpms2DMs. (A) Secondary sex traits were assessed to identify fish from each category. Representative images show body shape and color; insets are magnified views of the anal fin. (B) Adult sex ratios of progeny pooled from three triple heterozygote intercrosses for given genotypes. All double mutants also heterozygote for dmrt (MMH) were male as previously observed in rbpms2DMs, and all dmrt1 mutants in any other genetic combination (HHM, MHM, etc.) were female, as previously reported for dmrt1uc27/uc27. All triple mutants (MMM) were sterile males. Numbers indicate number of animals for each condition. Animals mutant for dmrt1 and any combination of a wild-type or heterozygous for a rbpms2 allele (WMM, WHM, HHM, etc.) were combined because they were phenotypically wild type, with the exception of rbpms2;dmrt1HMM as it had a different phenotype. The X2 goodness of fit test with Bonferroni correction was used for multiple comparisons. ***P<0.001 indicates groups that deviated from the expected 50:50 sex ratio. (C) Fertility was assessed in mating trials as indicated in the chart. For each cross, if a male triggered spawning or if sperm was acquired through IVF, the numbers (in parentheses) of fertilized eggs per trial for natural breeding are indicated. n/a indicates fish that were not able to be mated for subsequent mating trials. (D-G) Representative lateral view images of representative adult with tissue resected to reveal gonads (outlined in yellow dashed lines) of wild-type female (D), wild-type male (E), dmrt1uc27/uc27 (rbpms2;dmrt1HHM) female (F) and rbpms2;dmrt1MMM (G) fish. TM fish lacked morphologically obvious gonads proximal to the swim bladder (SB) compared with fertile sibling wild type or single mutants.