A pharmacohistochemical method for gamma-aminobutyric acid transaminase, the key enzyme for gamma-aminobutyric acid metabolism, has been combined with retrograde tracing by horseradish peroxidase, to a study of projections from the midbrain to the superior colliculus. The results indicate projections from the substantia nigra zona reticulata, the zona incerta and the reticular formation of the mesencephalon which are exclusively from neurons staining intensely for gamma-aminobutyric acid transaminase and presumptively gamma-aminobutyric acid as their transmitter. The projection from the ventral lateral geniculate body to the superior colliculus, on the other hand, comes from cells which do not stain for gamma-aminobutyric acid transaminase and therefore do probably not use gamma-aminobutyric acid as their transmitter.