Fig. 1

Zebrafish oocyte maturation mutants. A. Wild-type stage IV oocyte and egg in incident light. Eggs of p33bjta (1570 from 21 females), poacp26ahubb (1620 from 35 females), and p35aluc (2088 from 25 females) resemble wild-type oocytes in their opacity, while eggs of p26ahubg (546 from 8 females) and blacp25bdth (1020 from 17 females) were degenerated. B. Images of 5 minutes post-egg activation (mpa) wild-type (n = 68), p26ahubg (n = 56) and blacp25bdth (n = 61) eggs, showing lysed mutant eggs. bd, blastodisc. C. Non-complementation test showed that females carrying the ovyp37ad mutation in trans to the p35aluc mutation produced opaque mutant eggs (n = 1382 from 9 females), showing that p35aluc is a new ovy allele. D. Coomassie stained gel of major yolk proteins in wild-type and mutant eggs showing that the cleavage of yolk protein is impaired. kD, kilodaltons. E. Low (top row) and high (bottom row) magnification histological sections of wild-type and mutant stage III oocytes stained with H&E revealing YG sizes. Scale bar = 96 μm (top row) and 16 μm (bottom row). F. Scatter plots of YG diameters measured in a defined region of the oocyte. The mean diameter of the YGs in the mutants was significantly different to that of wild-type oocytes (WT = 14.6 μm, p33bjta = 3.87 μm, p35aluc = 5.95 μm, p25bdth = 9.25 μm). The number of YGs analyzed in this study was: wild type (n = 188 from 4 oocytes), p33bjta (n = 320 from 2 oocytes), ovyp35aluc (n = 503 from 2 oocytes), and blacp25bdth (n = 177 from 2 oocytes). Data are means ± SEM. ****p<0.0001 and **p = 0.0066 in parametric statistical Tukey’s test. ns, not significant. G. Confocal z-projections of isolated acid fixed oocytes analyzed by fluorescent in situ hybridization and also DiOC6 staining (green) in lower panels. In stage II oocytes, cycB1 transcript localized to the animal pole in wild-type (n = 32), p33bjta (n = 22), poacp26ahub (n = 47), ovyp35aluc (n = 38), and blacp25bdth (n = 28) oocytes. In stage I oocytes, dazl mRNA co-localized with DiOC6 to the vegetal Balbiani body (Bb) in wild-type (n = 32), p33bjta (n = 23), poacp26ahub (n = 63), ovyp35aluc (n = 31), and blacp25bdth (n = 29) oocytes. Scale bar = 35 μm.