Chronic antidepressant treatment has been shown to increase adenylyl cyclase activity, in part, due to translocation of G␣ s from lipid rafts to a nonraft fraction of the plasma membrane where they engage in a more facile stimulation of adenylyl cyclase. This effect holds for multiple classes of antidepressants, and for serotonin uptake inhibitors, it occurs in the absence of the serotonin transporter. In the present study, we examined the change in the amount of G␣ s in lipid raft and whole cell lysate after exposing C6 cells to escitalopram. The results showed that chronic (but not acute) escitalopram decreased the content of G␣ s in lipid rafts, whereas there was no change in overall G␣ s content. These effects were drug doseand exposure time-dependent. Although R-citalopram has been reported to antagonize some effects of escitalopram, this compound was without effect on G␣ s localization in lipid rafts, and R-citalopram did not inhibit these actions of escitalopram. Escitalopram treatment increased cAMP accumulation, and this seemed due to increased coupling between G␣ s and adenylyl cyclase. Thus, escitalopram is potent, rapid and efficacious in translocating G␣ s from lipid rafts, and this effect seems to occur independently of 5-hydroxytryptamine transporters. Our results suggest that, although antidepressants display distinct affinities for well identified targets (e.g., monoamine transporters), several presynaptic and postsynaptic molecules are probably modified during chronic antidepressant treatment, and these additional targets may be required for clinical efficacy of these drugs.The selective serotonin reuptake inhibitors (SSRIs) are the most frequently used drugs for treatment of depressive and anxiety disorders. However, the mechanism of action at the molecular and cellular level remains unclear. Certainly, one effect of SSRIs is binding to the 5-HT transporter and preventing reuptake of 5-HT into serotonergic neurons, increasing serotonergic transmission. However, 5-HT reuptake inhibition occurs rapidly, whereas the clinical effects require several weeks of drug administration. So, gradual changes should happen in the brain that can match the delayed response of antidepressants. There are several actions of SSRIs that are of delayed onset. SSRIs seem to elicit hippocampal neurogenesis (Santarelli et al., 2003); however, the linkage of this phenomenon to behavior responses to antidepressants has not been established (Wang et al., 2008). Synaptic rearrangement has also been associated with antidepressant action (Guest et al., 2004), and this is often linked to increased brain-derived neurotrophic factor, which, in turn, results from up-regulated cAMP signaling (Malberg and Blendy, 2005;Gass and Riva, 2007).On the molecular level, it has long been established that chronic antidepressant treatment increases the coupling between G␣ s and adenylyl cyclase. Menkes et al. (1983) found that long-term antidepressant treatment enhanced guanylyl-5Ј-imidodiphoshate [Gpp(NH)p]-and fluoride-stimulated a...