We investigated whether direct, cortico-motoneuronal connections are present in the rat, using both light microscopic and electron microscopic techniques. Corticospinal fibres were labelled using the anterograde tracer, biotinylated dextran-amine (BDA), which was injected into forelimb sensorimotor cortex. Motoneurons were retrogradely labelled after injection of cholera toxin subunit B (CTB) into forelimb muscles, contralateral to the injected hemisphere. Terminals of peripheral afferent fibres, which were also labelled by CTB, were easily distinguishable from, and much larger than, BDA-labelled corticospinal terminals. At the light microscope level, corticospinal terminals were found in all laminae contralateral to the injection site, most extensively in laminae VI and VII of cervical segments C5-C8. Although labelling in the ventral horn (lamina IX) was present, it was extremely sparse. A total of 47 corticospinal synapses were studied at the electron microscope level; most of these were in lamina VII and the majority (35/47; 74%) made axo-dendritic contacts with asymmetrical synapses; one made an axo-somatic synapse, and in the remaining 11 cases no postsynaptic structure could be identified. All corticospinal terminals contained spherical boutons. Serial sectioning of eight BDA-labelled corticospinal boutons in lamina IX revealed that most (seven out of eight) did not make synaptic contacts with any neuronal structure, and none made any contact with adjacent dendrites of CTB-labelled motoneurons. Thus these results provide no positive ultrastructural evidence for direct cortico-motoneuronal synaptic connections within lamina IX between corticospinal axon boutons and the proximal dendrites of forelimb motoneurons. The results confirm other lines of evidence suggesting that such connections are not present in the rat.