Abstract It is well documented that acute stress activates the sympathetic nervous system (SNS) and the Hypothalamus–Pituitary–Adrenal (HPA) axis. Results regarding the hypothalamus pituitary gonadal (HPG) axis, in contrast, are less consistent. Stress-associated increases as well as decreases have been reported for testosterone and estradiol. In the present study, healthy young male (n = 39) and female participants (n = 44, all tested in the luteal phase) were randomly assigned to a well-evaluated psychosocial stress protocol (“Trier Social Stress Test”, TSST) or to a non-stressful control condition (“Placebo-TSST”). Salivary concentrations of cortisol, alpha-amylase, testosterone, progesterone, and estradiol were measured immediately before and twice (10 and 25 min) after the treatment. As was to be expected, cortisol- and sAA-concentrations increased in response to the stressor. Stressed men showed a more pronounced increase of cortisol than stressed women. In contrast, acute stress did not affect testosterone-, progesterone-, and estradiol-concentrations. The results of the present study suggest that an acute psychosocial laboratory stress or has no strong rapid effects on salivary gonadal steroids. In line with several previous studies the findings might suggest that stress-induced changes in gonadal steroids occur in response to physical stressors, to competitive stressors or to more severe stressors only.