Sex differences and estrous cycle variations in anxiolytic-like behaviors and progestin concentrations were examined. Proestrous (n = 22), estrous (n = 19), diestrous (n = 20), and male (n = 18) Long-Evans rats were tested in horizontal crossing, open field, elevated plus-maze, emergence, holeboard, social interaction, tailflick, pawlick, and defensive burying tasks. Concentrations of plasma and hippocampal progesterone and 5 alpha -pregnan-3 alpha -ol-20-one (3 alpha ,5 alpha -THP) were measured by radioimmunoassay in behaviorally tested (proestrus n = 11, estrus n = 8, diestrus n = 9, male n = 7) and yoked non-tested rats (proestrus n = 11, estrus n = 8, diestrus n = 10, male n = 8). Proestrous females exhibited mole anxiolytic-like behavior than all other groups on the elevated plus-maze, social interaction, and defensive burying tasks. Proestrous females had significantly shorter latencies to emerge from a cylinder than did estrous and diestrous females, but not males. Proestrous and estrous females entered significantly more peripheral and total squares in a brightly-lit open field than did males. While proestrous females had a tendency to make more beam breaks than did males in the horizontal crossing task, there were no differences between groups on the holeboard task. There was a tendency for proestrous females to have longer tailflick latencies than diestrous and male rats; however, on the pawlick task there were no differences among the groups. Plasma and central progesterone and 3 alpha ,5 alpha -THP of tested and non-tested rats were not different. Proestrous females had significantly higher plasma and hippocampal progesterone and 3 alpha ,5 alpha -THP levels than all other groups. These data demonstrate that proestrous increases in anxiolytic-like behavior coincide with elevated circulating and hippocampal progestin concentrations. (C) 2000 Elsevier Science Inc. All rights reserved.