BackgroundThe serotonergic neurotransmitter system is involved in many ethanol‐induced changes, including many behavioural alterations, as well as contributing to alcohol dependence and its withdrawal.AimsThis review has evaluated microdialysis studies where alterations in the serotonin system, that is, serotonin, 5‐HT, or its metabolite 5‐hydroxyindoleacetic acid, 5‐HIAA, have been reported during different ethanol intoxication states, as well as in animals showing alcohol preference or not. Changes in 5‐HT receptors and the 5‐HT transporter are briefly reviewed to comprehend the significance of changes in microdialysate 5‐HT concentrations.Materials and methodsChanges in 5‐HT content following acute, chronic and during ethanol withdrawal states are evaluated. In addition, the serotoninergic system was assessed in animals that have been genetically selected for alcohol preference to ascertain whether changes in this monoamine microdialysate content may contribute to alcohol preference.Results and discussionChanges occurred in 5‐HT signalling in the limbic brain regions, increasing after acute ethanol administration in specific brain regions, particularly at higher doses, while chronic alcohol exposure essentially decreased serotonergic transmission. Such changes may play a pivotal role in emotion‐driven craving and relapse. Depending on the dosage, mode of administration and consumption rate, ethanol affects specific brain regions in different ways, enhancing or reducing 5‐HT microdialysate content, thereby inducing behavioural and cognitive functions and enhancing ethanol consumption.ConclusionMicrodialysis studies demonstrated that ethanol induces several alterations in 5‐HT content as well as its metabolites, 5‐HIAA and 5‐HTOL, not only in its release from a specific brain region but also in the modifications of its different receptor subtypes and its transporter.