Figure 4. Mutation of ndrg1b alters the gametogenesis and reproduction success of both sexes in adults.
Schematic representation of the experimental procedure adopted to analyze the effect of ndrg1b loss of function in the reproductive success at 80 days post hatching (dph) (A). Reproductive success was evaluated by crossing the wild-type (wt) males with sgN1b females (cyan full), wt females with sgN1b males (cyan empty), and as a control, wt females with wt males (gray). For all crossings, we quantified the total number of eggs spawned (B), percentage of fertilization (C), and percentage of hatching (D) of the total eggs spawned. The biallelic mutants of ndrg1b did not exhibit morphological sexual differences: adult females with their ovaries, wt (E) and sgN1b (G), and adult males with their testis, wt (F) and sgN1b (H). The secondary sexual characteristics are indicated by the separation of hindmost rays from other rays in the dorsal fin. Scale bars are 1 mm. Histological transverse sections of wt and sgN1b gonads from XX (I) or male (J) individuals. In the testis, each cyst of germ cells (spermatogonia-like cells) is encircled by a black dotted line. Scale bars represent 500 µm (I) and 50 µm (J). Number of oogonia (O) (K), number of pre-vitellogenic oocytes (PVO) (L), number of vitellogenic oocytes (VO) (M), number of spermatogonial cysts (N), number of spermatogonia per cyst (O), and number of spermatozoa (P) from wt female or male individuals (represented by gray empty bars or full bars, respectively) and sgN1b female or male individuals (represented by cyan empty bars or full bars, respectively). n = 10 per group. p-Values are indicated when groups differ significantly (p<0.05). NS: not statistically significant. Tukey´s multiple comparisons test (B–D). Unpaired Student’s t-test (K–P).
Figure 4—figure supplement 1. F1 reproductive success was evaluated by crossing the wild-type (wt) males with F1 females (cyan full), wt females with F1 males (cyan empty), and as a control, wt females with wt males (gray).
