We reported previously that substance P (SP)-containing projection neurons (SP-PN) in the striatum emitted many axon collaterals within the striatum (J. Comp. Neurol. 388 (1997) 250), and that substantially all striatal interneurons showing immunoreactivity for nitric oxide synthase (NOS: synthetic enzyme for the freely-diffusible messenger nitric oxide) displayed immunoreactivity for SP receptor (NK1: NK-1-type tachykinin receptor; Neurosci. Lett. 310 (2001) 109). By combining immunohistochemistry for NOS with immunogold labeling for SP, the present study revealed that SP-immunoreactive axon terminals were in synaptic contact with NOS-immunoreactive aspiny neurons in the rat striatum, indicating that SP-PN in the striatum sent their axon collaterals to nitric oxide synthase-expressing interneurons (NOS-IN) in the striatum. On the basis of these present and previous data, possible synaptic and non-synaptic interactions between SP-PN and NOS-IN in the striatum were discussed.