Restraint represents psychological and physical stress. Methods used to model restraint stress in mice vary in duration, time of day during which restraint is applied, and the strain of mouse tested. The goals of this study were: (1) to identify the optimal daily time periods during which the stress response is maximized, and (2) to describe mouse strain differences, if any, in response to restraint. Groups of outbred CD-1 and inbred C57BL/6 mice were restrained for 3 h during three time points of the daily light-dark cycle: (1) the late light phase, (2) the transition between the light phase and the dark phase, and (3) the mid-dark phase. Additional mice served as control groups for food deprivation or were unhandled except for blood sampling. Mice of both strains lost significant body mass after 3 days of restraint. Unrestrained food-deprived mice lost body mass, particularly if food-deprived during transition periods. Corticosterone was elevated in restrained mice compared with control mice. Neither basal nor postrestraint corticosterone differed between strains. Corticosterone was elevated by food deprivation during transitional periods in CD-1 mice and during both transition and dark phases in C57 mice. Corticosterone response in restrained CD-1 mice was increased during the dark phase. These results suggest that the physiological response to restraint is similar in both strains. However, corticosterone responses to both restraint and food deprivation were highest during the transitional and dark phases. (C) 2003 Elsevier Inc. All rights reserved.