Dorsal horn spinocerebellar tract cells of adult rats were labelled by retrograde axonal transport with the B subunit of cholera toxin. Sections were prepared from lumbar and thoracic spinal segments and incubated with antisera which specifically recognise neurokinin-1 receptor protein and substance P. Labelled cells and immunoreactivity for the receptor and substance P were identified by using three different fluorophores and the relationships between them were assessed in single optical sections with three-colour confocal laser scanning microscopy. Forty-eight cells were examined and 23 of them displayed immunoreactivity for the receptor. Many substance P-immunoreactive profiles were present in lamina V and some formed contacts with spinocerebellar tract cells possessing neurokinin-1 receptor immunoreactivity. The evidence suggests that substance P may influence the activity of a subpopulation of dorsal horn spinocerebellar tract cells by acting through neurokinin-1 receptors.