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Fig. 5
Wnt Signaling Is Activated and Regulates Ovary Regeneration
(A) RNA fluorescent in situ hybridization of lef1 shows upregulation of lef1 expression at 2 dpa compared to 0 dpa.
(B) lef1 expression level measured by qRT-PCR (∗∗∗p < 0.001, two-tailed t test, error bars indicate SEM).
(C) The ovary regeneration in dkk1b-overexpressing fish is blocked at 5 dpa compared to heat-shocked wild-type (hsWT). The red lines represented the distance between two remaining ovarian tissues after amputation.
(D) Quantification of the red line lengths of hsWT and hsDkk1b at 0 dpa and 5 dpa for 7 samples in the same group (n = 7).
(E) Quantification of regenerative ratio of hsWT and hsDkk1b at 5 dpa (n = 7, mean ± SEM, ∗∗∗p < 0.001, two-tailed t test, error bars indicate SEM).
(F) Triple fluorescent labeling with nanos2 RNA probe, Vasa antibodies, and DAPI staining both in the hsWT and hsDkk1b fishes at 5 dpa. Arrowheads point to GSCs.
(G) Quantification of the number of GSCs in the ovarian remnant per left ovary at 5 dpa (n = 7, mean ± SEM, ∗∗p < 0.01, two-tailed t test, error bars indicate SEM).
(H) nanos2 expression level measured by qRT-PCR (n = 7, mean ± SEM, ∗∗∗p < 0.001, two-tailed t test, error bars indicate SEM).
Scale bars: 100 μm (A), 1 mm (C), 50 μm (F). See also Figures S4 and S5.
(F–F″) Testis (arrowheads) that lack germ cells appears at 90 dpt, showing that the females treated with Mtz reverted to sterile males.
Oo, oogonia; IAz, zygotene-stage-IA; IA, stage IA oocytes; IB, stage IB oocyte; II, stage II oocyte; III, stage III oocyte; IV, stage IV oocyte; V, stage V oocyte. Scale bars: 50 μm (A), 1 mm (B–F′), 200 μm (B″–F″). See also Figure S2.