The ability of immobilization stress (IMO) to decrease Leydig cell steroidogenesis and serum androgen level is well documented, but the possible mechanism(s) involved in the adaptation to prolonged or repeated stress have not been identified. In this study we investigate impact of two sister kinases, protein kinase A (PKA) and protein kinase G-I (PKG-I) in stress adaptive responses of Leydig cells obtained from adult rats subjected to acute (2h) and repeated IMO (2 or 10 days, 2 hours daily). Results showed reduced cAMP and unchanged cGMP production of Leydig cells isolated from animals exposed to both acute and repeated IMO. Despite the reduced cAMP production, immunoblot analysis revealed increased level of PKA as well as level and phosphorylation of mature steroidogenic acute regulatory (StAR) protein. In this scenario, increased level of PKG-I transcript and protein was observed. Also, repeated IMO increased level of StAR and PKG-I immunoprecipitated complex, and this complex was further enhanced with ex vivo soluble guanylate cyclase (sGC) stimulation, but reduced with sGC inhibition. Aditionaly, ex vivo treatment of Leydig cells isolated from control and IMO rats with specific PKA and PKG inhibitor (H89 and KT5823 respectivily) showed that repeated IMO-induced increase in StAR immunoreactivity was decreased by inhibition of PKG and PKA. Moreover, it looks like two inhibitors have an additive effect on decreasing of StAR protein level, as well as androgen production. Results presented here suggest significant impact of both PKA and PKG-I cascade dependent activation of StAR protein, as adaptive mechanism, in recovering of stress-disturbed Leydig cells androgen production.