Unveiling Distinct Developmental and Cellular Impacts of Cisplatin on Zebrafish (Danio rerio) Embryos: Application of a Combined Developmental Toxicity with Rapid DNA-Content Flow Cytometry Protocol.

Rapid cellular assessment of cell-cycle perturbation can complement conventional developmental toxicology in zebrafish, but rapid DNA-content workflows developed for mammalian tissues require adaptation for whole embryos. Here, we applied an embryo-adapted version of rapid DNA-content flow cytometry analysis within a 96-h cisplatin exposure model in zebrafish (Danio rerio) and integrated this cellular readout with morphometric, cardiac, and neurobehavioral analysis. Dechorionated embryos were exposed to 10-400 μM cisplatin and evaluated for body length, eye surface area, heart rate, locomotor activity, thigmotaxis, touch-evoked and vibrational startle responses, and whole-embryo cell-cycle distribution. The adapted workflow combines immediate mechanical dissociation, filtration, brief propidium iodide/RNase staining, singlet gating, and direct flow cytometric acquisition. Importantly, each flow cytometry sample consisted of a single embryo, enabling DNA-content analysis at the individual-embryo level without pooling. Cisplatin produced time- and concentration-dependent growth effects and high-dose behavioral alterations, whereas heart rate was comparatively preserved. Flow cytometry detected the clearest cell-cycle redistribution at 400 μM, with a significant reduction in G0/G1 and an increase in G2/M. These findings demonstrate the practical applicability of a combined rapid embryo-adapted DNA-content workflow as a complementary cellular endpoint in a zebrafish developmental-toxicity platform and support further methodological application across experimental settings.