Fig. 3
Schematic of the experimental design, with timelines and endpoints of the study. Fertility impairment was induced in the female zebrafish using only ENU mutagenesis, while in male zebrafish, ENU mutagenesis was followed by treatment with metronidazole. Mutagenized zebrafish producing less than 20 embryos were considered as fertility impaired models and used in the study. Various study groups comprising of healthy control, fertility impaired models, zebrafish treated with letrozole (female)/clomiphene (male), and zebrafish treated with six different doses of SLSO were used in the study (Fig. 4). After 14 days of dosing, the fish were allowed to spawn for 1 day, followed by data collection at endpoints 1 and 2 in females and males, respectively. In females, the ovulation cycle was allowed for the next 5 days, followed by dissection at endpoint 4 to assess the ovarian reserve of eggs. In a parallel experiment, female fish were dosed for 14 days and then dissected on the 15th day to study ovarian morphology, cytology, and cyst at endpoint 3