Sugammadex

The embryos were cultured in an incubator (Daihan Scientific, Wonju, Korea) at 28.0°C ± 1°C. To improve the permeability of the drug, a small hole was drilled and processed using forceps in the amnion of all embryos, except those in the negative control group. Sugammadex (MSD, Seoul, Korea) solutions of varying concentrations (0 [control], 25, 50, 75, 100, and 200 μg/mL) were prepared using E3 medium. About 1–2 hours post-fertilization (hpf), the embryos were randomly transferred into the Sugammadex solution in 12-well plates until 120 hpf. The exposure solution was refreshed every 24 hours to maintain the appropriate concentration of Sugammadex and quality of the E3 medium. The number of dead embryos and the state of development were observed every day. The dead embryos were removed when changing the exposure solution. Morphological development of the embryos was observed under a microscope. The larvae hatched 72 hpf, and body length was measured 96 hpf. The lethality was measured 120 hpf. The hatching and malformation of embryos were checked daily before changing the exposure solution. The body length and heart shapes were measured using a microscope (Nikon SMZ18, Tokyo, Japan). All experiments were performed in triplicates.