Figure 2 ∣.

Gestational Vitamin C deficiency has a long-term impact on female fecundity.

a, Diagram of experiments to address female fecundity (see Fig. 1a). From E13.5 onwards, Vitamin C was resupplied in the drinking water. After birth, F1 females were assessed for ovarian reserve at day 7 or fecundity at 20-25 weeks. All -VitC and control mice are of the same genotype, Gulo^−/−;Oct4/EGFP.

b, Representative whole-mount images of P7 ovaries from -VitC or control females. Oocyte nuclei identified by Nobox (red) are segmented and sorted by volume to quantify primordial and primary follicles. n = 9 ovaries per condition.

c, Quantification of oocytes in day 7 ovaries by whole-mount imaging of Nobox staining. Geometric shapes represent individual female embryos; Identical shapes indicate ovary pairs. Control females contain 3000-4500 Nobox+ oocytes/ovary (n=9). -VitC females contain 1000-4500 Nobox+ oocytes/ovary (n=9). Statistical significance assessed by two-tailed Student’s t-test and variance determined by F ratio. Centre line represents mean.

d, Normal numbers and variance in P7 ovary volume from females that had developed in the absence of Vitamin C. n = 9 ovaries per condition were measured, matched with data in Fig. 2c. Statistical significance assessed by two-tailed Student’s t-test and variance determined by F ratio. Centre line represents mean.

e, -VitC or control F1 females were mated to wild-type males, and the number of implantation sites per female displaying a vaginal plug was counted between E9.5-E18.5. Ctrl F1 females average 6.5 implants per pregnancy while -VitC F1 females average 4 implants. Statistical significance assessed by two-tailed Student’s t-test. Error bars depict mean ± SEM.

f, There is a significant reduction in the number of successful matings in -VitC F1 females. A successful mating is defined as the observation of at least 1 implanted embryo per female that had displayed a vaginal plug after mating. Data collected from n=14 Vitamin C deficient and n=17 control mated females. Statistical significance assessed by Chi-square test.

g, Within successful matings only, there is a higher number of resorptions in -VitC F1 females, relative to controls. Statistical significance assessed by two-tailed Student’s t-test. Error bars depict mean ± SEM.

h, The number of pregnancies containing at least one resorbed embryo is significantly higher in - VitC F1 females, relative to controls. Data collected from n=5 -VitC and n=12 control pregnant females. Statistical significance assessed by Chi-square test.