Introduction: We previously demonstred that vitamin A (retinol) induces reactive oxigen species (ROS) production and oxidative stress in cultured Sertoli cells.  This leads to an increase in proliferative indexes and cellular preneoplastic transformation, through the oxidant-dependent activation of the Src-MEK-ERK1/2-CREB signaling pathway. The AP-1 transcription factor is a heterodimer composed by elements of the Jun and Fos families and binds to DNA at specific AP-1 binding sites, and its activity is determined in part by phosphorylation of these complex components or by cystein specific oxidation. Since the AP-1 has been observed to be important in several mitogenic signaling pathways, we hypothesized that retinol-induced oxidative stress can modulate AP-1 signaling. Thus, in this work we investigated the effect of retinol treatment on AP-1 pathway modulation. Methods: Cultured Sertoli cells isolated from 15 days old Wistar rats were treated for different times with ranging concentrations of retinol (0,2-10  mM). Nuclear proteins were isolated by differential centrifugation of the nucleus followed by selective membrane rupture. Electro Mobility Shift Assay (EMSA) using oligonucleotides with AP-1 binding sequences was performed to avaliate AP-1 activation in nuclear extracts. Results: We observed that retinol treatment strongly suppressed AP-1 basal transcriptional activity in a concentration-dependent manner. We verified that antioxidant treatment with Trolox (100  mM) and N-acetylcysteine (1 mM) reversed the effects of retinol, which indicates that the cellular redox status is important to AP-1 transcriptional activity in this cells. These results suggest that some oxidative-triggered responses caused by retinol in Sertoli cells are mediated, at least in part, by suppression of AP-1 regulated genes.