Different technical, physiological and biochemical aspects concerning the study of the release of 5-HT are discussed herein. Isotopic methods are the most suitable techniques since these allow the release of 3H-5-HT to be measured after having determined the identity of the labelled compounds formed from 3H-tryptophan by co-chromatography. Under these conditions, the 3H-amine released in the superfusates comes from serotoninergic nerve endings, since tryptophan hydroxylase is exclusively localized in serotoninergic neurons. Moreover, it appears that newly synthesized 5-HT is preferentially released. The release of 5-HT is dependent on neuronal activity, but is not always linked to the synthesis of 5-HT. The increase in the firing rate of serotoninergic cell bodies by a local application of glutamate in the area of the nucleus raphe dorsalis induces a marked increase n the release of 5-HT in the caudate nucleus; an opposite effect is observed after cooling this region. The local depolarization of serotoninergic terminals located in the caudate nucleus increases the release of this amine. This effect is blocked by TTX. LSD reduces the stimulating effect of KCl, thus indicating that the release of 5-HT can be controlled at a presynaptic level. In addition, the release of the amine is dependent on the presence of calcium. Serotoninergic neuronal activity can be controlled at the preterminal or at the cell body levels by the activity of other neuronal systems. The effects of the release of dopamine from dendrites, and that of GABA in the substantia nigra are reported herein. Furthermore, changes in the activity of the dopaminergic, gabaergic and serotoninergic systems innervating the nucleus raphe dorsalis modulate the release of 5-HT, measured both in the caudate nucleus and in the nucleus raphe magnus. Finally, it has been reported that the release of 5-HT can be estimated in the raphe nuclei dorsalis and magnus. It has been shown that the amounts of 3H-5-HT continuously formed from 3H-TRP and released in the nucleus raphe dorsalis are much greater than those estimated in the caudate nucleus or in the substantia nigra. Although the quantities of endogenous 5-HT measured in the nucleus raphe dorsalis are the highest in the brain, this structure presents only a few serotoninergic nerve endings. This raises the question of the origin of the 5-HT released in serotoninergic nuclei. A possible dendritic release of 5-HT is discussed.