Journal: International Journal of Molecular Sciences

Article Title: Chr23-miR-200s and Dmrt1 Control Sexually Dimorphic Trade-Off Between Reproduction and Growth in Zebrafish

doi: 10.3390/ijms26041785

Figure Lengend Snippet: Dmrt1 regulates gonad development in males but not in females. ( A ) Anatomical observation of testes in WT and dmrt1 −/− at 5 mpf. The black and blue arrowheads indicate the testes in WT and dmrt1 −/− zebrafish, respectively. ( B ) Histological examination of testes in 5-month-old WT and dmrt1 −/− zebrafish. Transverse section of the abdomen by HE staining. The black and blue arrowheads indicate the testes in WT and the severely regressed testicular tissue in dmrt1 −/− zebrafish, while the blue dotted lines show the amplified images of testes in dmrt1 −/− zebrafish. Scale bars, 1 mm. ( C ) Immunofluorescence staining of WT and dmrt1 −/− testis transverse sections with Ddx4 antibody (green). The nuclei were stained by DAPI (blue). The white dotted lines show the testes in dmrt1 −/− zebrafish. Scale bars, 50 μm. ( D ) Anatomical observation of ovary in 5-month-old WT and dmrt1 −/− zebrafish. ( E ) Successful spawning rate of WT and dmrt1 −/− females mating naturally with WT males at 5 mpf. There were three independent repetitions ( n = 15). ( F ) Fertilization rate of WT and dmrt1 −/− females by crossing with WT males at 5 mpf ( n = 15). Data are represented as mean ± SD (ns, no significant).

Article Snippet: For immunofluorescence staining, the sections after deparaffinization were rehydrated in PBS, repaired in bath of 0.01 M citric acid antigen retrieval buffer (pH = 6) at 97 °C for 10 min. Then, slides were blocked with 5% BSA in PBS for 30 min at room temperature, and incubated overnight at 4 °C with rabbit polyclonal anti-Ddx4 (Vasa) primary antibody (1:100, #A15624; ABclonal, Wuhan, China).

Techniques: Staining, Amplification, Immunofluorescence