Fig. 5: Dmrt1 deficiency led to all female development in Juji. | Communications Biology

Fig. 5: Dmrt1 deficiency led to all female development in Juji.

From: Cyprinid Juji (Gobiocypris rarus) as a model fish to study germ cell development and gonadal differentiation

Fig. 5

A Schematic of dmrt1 gene structure with 5 exons and CRISPR/Cas9-induced mutation. Underlined sequence is CRISPR site, and protospacer adjacent motif (PAM) sequence is shown in red. Mutant line with an 8 bp deletion was generated for phenotype analysis. B Schematic of wild-type and mutant Dmrt1 protein structures. DM, DM DNA binding domain; Dmrt1, Double-sex mab3 related transcription factor 1 domain. Arrowhead points to the mutation site. C Gross morphology of gonads in dmrt1+/+ females, dmrt1+/+ males and dmrt1-/- fish. Asterisks indicate gonads. dmrt1-/- testis was not found. Scale bars: 5 mm and 4 mm, respectively. D Sex ratio of dmrt1+/+, dmrt1+/- and dmrt1-/- fish after maturation (n = 26 dmrt1+/+, n = 51 dmrt1+/-, n = 25 dmrt1-/-). All dmrt1-/- fish developed into females. E Fertility analysis of female adult fish of different genotypes crossed with WT males (n = 3 for each group, one-way ANOVA exhibited no significant difference, mean ± SEM). The dmrt1-/- females exhibited comparable fertility to WT and dmrt1+/- females. F Fecundity analysis of female adult fish of different genotypes crossed with WT males (n = 3 for each group, one-way ANOVA exhibited no significant difference, mean ± SEM). G Fluorescence intensity distribution in dmrt1+/- and dmrt1-/- mutants at 20 dpf (n = 20 for each group, unpaired two-tailed Student′s t-test; ns: Not significant). H Gonadal EGFP fluorescence in dmrt1-/- mutants at 30 dpf, 60 dpf, and 90 dpf. Scale bars: 1 mm, 2 mm, and 2 mm, respectively. I Fluorescence intensity of Bright and Faint groups in dmrt1-/- mutants at 90 dpf (n = 5 for each group, unpaired two-tailed Student′s t-test, mean ± SEM; ns: Not significant). J Histological analysis of gonads in dmrt1-/- fish at different ages (30 dpf, n = 5; 60 dpf, n = 7; 90 dpf, n = 6; 120 dpf, n = 10; 150 dpf, n = 7). IA, stage IA oocyte; IB, stage IB oocyte; II, stage II oocyte; III, stage III oocyte; IV, stage IV oocyte; GSC, germline stem cell. Scale bars: 25 µm, 50 µm, and 200 µm, respectively.

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