Fig. 5

Cyp19a1a is required for oocyte maturation and maintenance of female fates, independently of antagonism of dmrt1. (A-D) Representative lateral view images show the external secondary sexual characteristics of d120 adults: cyp19a1a;dmrt1MH (n=7) male (A), cyp19a1a;dmrt1HM (n=7) female and cyp19a1a;dmrt1MM (n=5) mutants (C,D). Male MH fish (A) had a slender body shape and lacked a genital papilla (indicated by arrow). Female HM fish (B) had a round body shape with protruding abdomen and extended genital papilla. Similar to MH males, MMs (C,D) exhibited male secondary sex traits. (E-H) However, upon dissection, MH males (E) had testes, HM females (F) had ovaries that contained pre-vitellogenic (PV) early-vitellogenic (EV) and vitellogenic (V) stages of oocytes, whereas MMs (G,H) had gonads that were atypical in overall structure and contained oocytes that only progressed to the PV stage. (I-L) Additional representative brightfield and Vasa (green) overview images of cyp19a1a;dmrt1MH (I), cyp19a1a;dmrt1HM (J) and cyp19a1a;dmrt1MM (K,L) gonads. (M) Fertility was assessed by natural mating or by IVF extraction of sperm followed by fertilization of eggs. For each cross or if sperm was acquired through IVF, the numbers (in parentheses) of fertilized eggs per trial are indicated. (N) Adult sex ratios of progeny pooled from heterozygote intercross and double heterozygote crossed to a mutant;heterozygote (HH×MH) for given genotypes. All cyp19a1auc38 mutants that were also heterozygous for dmrt1 (MH) were males, all dmrt1uc27 mutants that were also heterozygous for cyp19a1auc38 (HM) were female. All cyp19a1auc38/uc38;dmrt1uc27/uc27 double mutants appeared male based on secondary sex traits, but female based on primary gonad sex and were classified as sterile because of the lack of mature gametes. Numbers indicate individual animals for each condition. 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. (O) Representative gonads from two d145 cyp19a1a;dmrt1MM fish (total, n=4) show an ovary containing PV oocytes and a gonad lacking oocytes. (P-R) Representative lateral view images show the external secondary sex traits of d145 bmp15;dmrt1 adults. The bmp15;dmrt1MH (n=7) (P) and bmp15;dmrt1HM (n=7) (Q) fish display normal secondary sex characteristics of males and females, respectively. MMs (n=4) (R) had a female body shape but lacked protruding genital papillae. (S-U) Upon dissection, MH males (S) had testes, HM females (T) had ovaries and MMs (U) had ovaries that contained PV oocytes.