Employing a combination of fluorescent retrograde double labeling and immunofluorescence histochemistry for substance P receptor (SPR), we examined the collateral projection from single SPR-like immunoreactive neurons in the medullary dorsal horn (caudal subnucleus of the spinal trigeminal nucleus) to bilateral parabrachial nuclei in the rat. After injection of fast blue (FB) or diamidino yellow (DY) into the right or left parabrachial nucleus, respectively, single-labeled FB or DY neurons and double-labeled FB/DY neurons were observed mainly bilaterally in laminae I and II of the medullary dorsal horn. Some of the single-labeled FB or DY and double-labeled FB/DY neurons showed SPR-like immunoreactivity, especially in lamina I. In lamina I of left medullary dorsal horn which is ipsilateral to the DY injection into the PBN, the percentages of double-labeled FB/DY neurons to the total number of FB- or DY-labeled neurons were 34.0% or 20.2%, triple-labeled FB/DY/SPR neurons to the total number of FB/DY double-labeled or SPR-like immunoreactive neurons were 22.0% or 2.4%, respectively. In lamina I of the right medullary dorsal horn which is ipsilateral to the FB injection into the PBN, the percentages of double-labeled FB/DY neurons to the total number of FB- or DY-labeled neurons were 12.9% or 59.3%, triple-labeled FB/DY/SPR neurons to the total number of double-labeled FB/DY or SPR-like immunoreactive neurons were 24. 6% or 3.9%, respectively. The results suggest that some of the single SPR expressing neurons in the medullary dorsal horn might be innervated by substance P containing primary afferent fibers and transmit sensory information diffusely to bilateral parabrachial nuclei by way of their axonal collaterals.