The susceptibility of mouse neural stem/progenitor cells (NSPCs) to heavy-metal cytotoxicity was assessed by measuring cell viability following exposure to heavy metal chlorides (ZnCl2, CdCl2, CuCl2, and HgCl2, respectively). We determined half-maximal inhibitory concentration (IC50) values, subcytotoxic doses, capacity for neural differentiation, and morphological features of glial fibrillary acidic protein (GFAP) network at the subcytotoxic doses of heavy metal ions. Experiments were performed using two protocols for the exposure at subcytotoxic doses of heavy metal ions; these protocols included simultaneous exposure with the induction of NSPC differentiation and sequential exposure after the induction for 1 week. Exposure to HgCl2 using both protocols reduced the ratio of neuronal NSPC differentiation. Although sequential exposure to CdCl2 reduced the size of GFAP network, simultaneous exposure did not induce any change. In conclusion, image analyses of the cytoskeletal morphology of NSPCs as a novel tool for assessing neurodevelopmental cytotoxicity enabled us to obtain new information about the localization of cytoskeletal proteins.

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