The effect of serotonin depletion (p-chlorophenylalanine pretreatment) on habituation of exploratory motor activity, and on cortical and hippocampal [3H]muscimol binding in vitro, was examined in rats. It appeared that the very strong decrease in serotonin concentration abolished motor habituation in the open field and decreased [3H]muscimol binding to cortical and hippocampal brain slices. The GABA(A) receptor down-regulation was due to a decrease in the apparent affinity of the radioligand for the receptors. p-Chlorophenylalanine-induced biochemical changes were selective and most probably secondary to serotonin depletion, as the serotonin synthesis inhibitor did not displace [3H]muscimol from its binding sites in neural membranes taken from the occipital cortex. It is concluded that there is a functional interaction between brain serotonin and GABA (gamma-aminobutyric acid) systems, both at behavioral and biochemical levels, that is involved in the motor activity habituation process.