Meiotic defects and failure of germ cell maintenance in Tdrd12-deficient zebrafish. (A) Normalized expression of the meiotic recombination-specific gene sycp3 (synaptonemal complex protein 3) in the developing gonads were examined in gonadal tissue samples in Tdrd12-deficient fish and their wild-type siblings at 13, 18, 25, and 35 dpf; (B) In situ cell death in the immature testis of tdrd12+/+ and tdrd12−/− at 35 dpf, with abundant signals (brown) in the Tdrd12-deficient testis. Normalized expression of the germ cell-specific genes dnd (C), vasa (D), and piwil1 (E) in the developing gonads were examined in gonadal tissue samples in Tdrd12-deficient fish and their wild-type siblings at 13, 18, 25, and 35 dpf. The tails of the larvae were collected for genotyping, while the rest of the body was used for total RNA isolation. The numbers of the examined fish for the assays at the 13, 18, 25, and 35 dpf stages for each genotype were 25, 20, 12, and 10, respectively. The experiments were performed with three biological repeats. * indicates the difference at p < 0.05 vs. wild type; ** indicates the significant difference at p < 0.01 vs. wild type. β-actin2 was selected as the most suitable and invariant reference gene for our samples from gapdh, β-actin2, and ef1a testing according to the published reports [33,34].
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