FSH is the main regulator of Sertoli cell function. Nevertheless, several other effectors such as catecholamines can also stimulate these cells through the adenylyl cyclase transduction pathway. However, the expression of beta adrenergic receptors in Sertoli cells is a subject of controversy. The aim of the present study was to determine if there are physiologically functional beta adrenergic receptors in Sertoli cells and to which subtype(s) they belong. In freshly isolated Sertoli cells, isoproterenol, a non selective beta-adrenergic agonist, was found to stimulate cAMP production and tissue-type plasminogen activator secretion. Specific transcripts for the beta1 and beta2, but not beta3, subtypes were detected by RT-PCR analysis. Beta2 transcripts were the form expressed predominantly in Sertoli cells. Binding experiments carried out on freshly isolated and on cytospined Sertoli cells indicated that in both conditions, [125I]iodocyanopindolol binding was inhibited by a non-selective and a 2 selective antagonist, whereas a beta1 selective antagonist had no effect. Scatchard analysis of beta2 specific inhibition revealed a dissociation constant of 0.3 nM and a receptor density of 14000 sites per cell. In freshly isolated Sertoli cells, we observed that cAMP and tissue-type plasminogen activator were stimulated by isoproterenol and a beta2 selective agonist, but not by beta1 or beta3 selective agonists. Accordingly, the isoproterenol-stimulated tissue-type plasminogen activator responses were abolished by the beta2 selective antagonist only. In cultured Sertoli cells, the trend was the same: tissue-type plasminogen activator and transferrin secretions were increased by isoproterenol and beta2 but not by beta1 or beta3 selective agonists. We conclude that freshly isolated Sertoli cells express beta2 adrenergic receptors which are functionally coupled to adenylyl cyclase and that these characteristics are preserved in cell culture. For the tested parameters, catecholamines and FSH effects were similar, but response magnitudes were systematically lower with beta agonists than with FSH. As norepinephrine is normally present in physiologically-relevant amounts in the interstitial fluid, it can be suspected to play a role in the regulation of Sertoli cell function.