In order to test the usefulness of the in vitro HRP-labeling technique in the brains of small lizards, the fasciculus retroflexus was labeled in isolated brains of young specimens of the lizard Gallotia galloti by means of HRP applied with the tip of a micropipette located approximately midway along the descending course of the tract. Cells in the medial and lateral habenular nuclei were labeled retrogradely. Anterograde transport showed the course of the fasciculus retroflexus, first dorsoventrally into the paramedian prerubral tegmentum and then longitudinally into the isthmic nucleus interpeduncularis. Some fibers spread dorsally into the mesencephalic and isthmic tegmental fields. A substantial bundle continues caudally into nucleus raphe parvocellularis. Other fibers diverge rostral-wards from the point of inflexion of the fasciculus, coursing longitudinally in the medial forebrain bundle into the basal telencephalon. Some fibers course rostrally from the nucleus habenularis lateralis in the stria medullaris and could be followed to the dorsal septum.