A NEW NEUROTOXICOLOGICAL TEST SYSTEM USING NERVE CELL LINE

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Many new chemicals have produced in this century from industrial circles. We established a in vitro neurotoxicological test system for the purpose of assessing the neurotoxic potential of these chemicals.(1) Induction of micronuclei and apoptosisHybrid cell line N18DRG-D2 of BALB/c mouse dorsal root ganglion neuron and mouse neuroblastoma cell (N18TG-D2) was used in this study. Induction of micronuclei by propidium iodide peaked at 96 hours of cisplatin treatment and frequency of micronuclei induced by cisplatin increased dose dependent manner. However, number of apoptotic micronuclei (double staining with proidium iodide and TUNEL method) did not increase following cisplatin treatment.(2) Alteration of membrane action potentialMembrane action potential was determined using a fluorophotometer after stained with 3, 3'-dipenty loxacarbocyanine iodide. Fluorescence intensity of N18DRG-D2 was attenuated after exposure to cisplatin.(3) Alteration of intracellular calcium ion concentrationMembrane action potential was determined using a fluorophotometer after stained with fura-2. Fluorescence intensity of N18DRG-D2 was attenuated after exposure to cisplatin.These results suggested that a new neurotoxicological test system using N18DRG-2 cell line might be useful for estimating neurotoxic substances.