Figure 3.

DHA promotes oocyte quality through production of pregnenolone

A: Representative images of anti-α-tubulin staining of mature follicles from WT, fat1, and elovl2^-/- zebrafish treated with methanol (Ctrl) or 1 μg/mL nocodazole. Scale bar: 50 μm. B: Quantification of α-tubulin intensity of groups in panel A. Relative α-tubulin intensity was calculated by comparing fluorescence area of embryos from each experimental group with that of the WT group treated with methanol per unit area. At least three embryos were measured per group. C: Pregnenolone (P5) content in WT, fat1, and elovl2^-/- embryos at 15 mpf. D, E: Representative images and quantitative analysis of anti-α-tubulin staining of WT, fat1, and elovl2^-/- embryos at 6 hpf. Scale bar: 25 μm. At least four embryos were measured per group. F: P5 content in WT, fat1, and elovl2^-/- embryos at 6 hpf. G, H: Representative images and quantitative analysis of anti-α-tubulin staining of elovl2^-/- embryos incubated with methanol (MeOH, Ctrl) or P5 for 6 hpf. Scale bar: 25 μm. I: Real-time qPCR of cyp11a1 expression in WT, fat1, and elovl2^-/- eggs and 6 hpf embryos. J–L: Representative images and statistical analysis of WT, fat1, and elovl2^-/- embryos with normal development or knockdown of cyp11a1 at 10 hpf. Scale bar: 1 mm. Approximately 300 embryos were measured per group. M, N: Representative images and quantitative analysis of anti-α-tubulin staining of cyp11a1 knockdown embryos at 6 hpf from WT, fat1, and elovl2^-/- zebrafish. Scale bar: 25 μm. At least four embryos were measured per group. O, P: Representative images and quantitative analysis of anti-α-tubulin staining of elovl2^-/- embryos with normal development or injected with cyp11a1 mRNA. Scale bar: 25 μm. At least three embryos were measured per group. All values are mean±SD. Student’s t-tests were used in panels C, E, F, H, L, N, P. Multiple t-tests-one per row were used in panels B, I. ^*: P<0.05; ^**: P<0.01; ^***: P<0.001.