Diazepam, administered to rats at a high dose (25 mg/kg PO) has been shown to have no effect on the plasma corticosterone response to the stress of an elevated open platform. It did however, reduce the plasma corticosterone in rats repeatedly exposed to the apparatus. Diazepam-withdrawal from stress-habituated rats increased plasma corticosterone (p less than 0.01) whereas withdrawal of diazepam from unstressed rats had no effect on plasma corticosterone. It is concluded that this effect of diazepam-withdrawal may reflect the development of dependence upon the drug. Significant effects were not observed following the administration of a lower non-selective dose (5 mg/kg PO) of diazepam and, therefore, it is not clear whether dependence to its sedative, rather than the anxiolytic properties have been measured. Acute diazepam (25 mg/kg) increased (p less than 0.05) hippocampal 5-hydroxyindoleacetic acid; its withdrawal from unstressed rats after 40 days reduced (p less than 0.01) hypothalamic 5-hydroxytryptamine. There was no evidence that the effects of diazepam or its withdrawal on plasma corticosterone in stressed rats were associated directly with changes in brain 5-hydroxyindoles.