Fig. 5

Mios promotes oogenesis through mTorc1 signaling.

a–d Immunostaining of p-Ps6k (purple) in 35 dpf wildtype (n = 7) and mios^-/- (n = 10) germ cells. Germ cells are labeled with Ddx4 (teal) and nuclei are labeled with DAPI (white). a, b Arrows indicate p-Ps6k localization in mitotic and early meiotic nuclei. c, d Dashed lines indicate early oocytes. Adjacent panel shows p-Ps6k and DAPI localization in indicated cells. Scale bars are 20 µM. e Quantification of the intensity of p-Ps6k normalized to the intensity of Ddx4 for wildtype (n = 21 cells from 7 individual fish) and mios^-/- (n = 25 cells from 8 individual fish). Two-tailed paired equal variance Student’s t tests were performed for the indicated groups. P = 0.05. fmTOR^ca transgene: the human mTOR^ca expression is driven by the germline ziwi promoter and the heart specific promoter cmlc2 drives mCherry expression as a selectable marker. Scheme depicts injection into 1-cell embryos and generation of stable Tg[ziwi:mTOR^ca] lines. g Sex ratios for 90 dpf+ fish with ms49 and ms64 mTOR^ca alleles. Pink represents female and blue represents males; numbers of fish screened are indicated for each group. Non-transgenic siblings are indicated by - and transgenic siblings are indicated with + and the specific mTOR^ca alleles are indicated. Statistical significance was evaluated by performing a two-tailed χ² test with Bonferroni correction against the mios +/+; +/- non-transgenic control group. P = 0.0125. h Fertility assays and quantifications of total eggs, degenerating eggs, and fertility rates for ms49 and ms64 Tg[ziwi:mTOR^ca] fish (n = 3 fish per genotype and transgenic condition). Non-transgenic siblings are indicated by - and transgenic siblings are indicated with + and the specific mTOR^ca alleles are indicated. Two-tailed paired equal variance Student’s t tests were performed for the indicated groups. P = 0.05. All other comparisons were non-significant. e, h For box and whisker plots, line indicates median, boxes indicate upper (75th) and lower quartiles (25th), whiskers indicate data within 1.5 times the inner quartile range, and error bars represent minimum and maximum values. Any data outside of this range are plotted as individual points. Source data are provided as a Source Data file.