Agonist-induced mu-opioid receptor (MOPr) internalization has long been suggested to contribute directly to functional receptor desensitization and opioid tolerance. In contrast, recent evidence suggests that opioid receptor internalization could in fact reduce opioid tolerance in vivo, but the mechanisms that are responsible for the internalization-mediated protection against opioid tolerance are controversely discussed. One prevailing hypothesis is, that receptor internalization leads to decreased receptor signaling and therefore to reduced associated compensatory changes in downstream signaling systems that are involved in the development of opioid tolerance. However, numerous studies have demonstrated that desensitized and internalized mu-opioid receptors are rapidly recycled to the cell surface in a reactivated state, thus counteracting receptor desensitization and opioid tolerance. Further studies revealed agonist-selective differences in the ability to induce opioid receptor internalization. Recently it has been demonstrated that the endocytotic efficacies of opioids are negatively correlated to the induced opioid tolerance. Thus, clearer understanding of the role of opioid receptor trafficking in the regulation of opioid tolerance and dependence will help in the treatment of patients suffering from chronic pain or drug dependence.