The relationship between neuronal function in the brain and neuropsychological behavior were analyzed in young rats fed a zinc-deficient diet for 1–2 weeks. Serum zinc concentration was less than 50% of that of the control. However, zinc concentration in the hippocampal perfusate measured by the in vivo microdialysis was not decreased after 2-week zinc deprivation. Timm's stain, with which histochemically reactive zinc in the presynaptic vesicle is detected, was not also attenuated in the brain. On the other hand, serum corticosterone concentration, which was determined in the morning, was markedly increased after 2-week zinc deprivation and intracellular calcium signal, which was determined by fura-2 AM, was also increased in the hippocampus. In the hippocampus in zinc deficiency, intracellular free calcium concentration may be altered prior to the decrease in zinc concentration in the extracellular fluid. When rats were subjected to the open-field test, the frequency of line crossing and the time of grooming were decreased after 2-week zinc deprivation. In the plus-maze test, the time spent in the open arms was also decreased in zinc-deficient rats, suggesting that anxiety-like behavior is increased in zinc deficiency. The present study indicates that the increase in anxiety-like behavior in zinc deficiency may be linked to the increased concentration of basal free calcium in hippocampal cells, probably due to the increase in serum corticosterone concentration.