Pre-embedding immunoperoxidase (for serotonin) and postembedding immunogold (for γ-aminobutyric acid; GABA) labelling were combined at light and electron microscopic levels to demonstrate the neuronal targets of serotonin (5-HT) afferents in the ventral posterior lateral nucleus (VPL) of the cat thalamus. 5-HT-immunoreactive fibres and terminal varicosities were found in close proximity to GABA-immunoreactive interneurons and non-GABAergic relay neurons. Ultrastructurally, the vast majority of 5-HT terminals made close membrane contacts without overt membrane specializations with GABAergic axon terminals, GABAergic presynaptic dendrites and GABAergic somata. A very small number of 5-HT terminals formed typical asymmetrical synapses with GABAergic presynaptic dendrites and with dendritic shafts of relay cells. Some 5-HT terminals participated with the presynaptic dendrites in triadic synaptic arrangements. These findings suggest a dual innervation pattern by 5-HT afferents in VPL and the release of 5-HT in large part at sites not associated with morphologically detectable synapses.