Evaluation of marble burying behavior (MBB) housing-induced alteration of monoamine metabolism in mouse brain was performed by measuring metabolite levels with HPLC-ECD. Isolated housing of mice, each in a cage (22 x 32 x 14 cm; sawdust, 1-mm diameter; 5 cm in thickness) with 15 evenly spaced glass marbles on the floor (2.5-cm diameter; control, without marbles) for 24-168 hr, 5-HIAA contents were decreased in three regions: the midbrain, thalamus and hypothalamus. 5-HT turnover was not inhibited except for in the hypothalamus due to the decreases of 5-HT in the other two regions. On the other hand, DOPAC content and DA turnover were decreased in four regions: striatum, midbrain, thalamus, hypothalamus. The decrease in hypothalamic monoamine neurons was observed notably after 72 hr of MBB housing. No alterations were observed in feeding, water-drinking, spontaneous locomotor activity, number of buried marbles, serum corticosterone and serum glucose concentrations during MBB keeping compared with the control mice. These results suggested that the isolated mouse housed in a cage with evenly spaced glass marbles for a long period may be a model animal for alteration of monoamine metabolism in brain regions without physical infringement.