Time-dependent changes in Bolton-Hunter-labeled 125I-substance P binding occurred in the dorsal horn of the spinal cord following unilateral adjuvant-induced inflammation in the hindpaw of the rat. Inflammation was characterized by measures of edema and hyperalgesia. Edema and hyperalgesia were both present 6 h after induction of inflammation. However, by eight days, hyperalgesia had dissipated while edema persisted. Six hours after the induction of inflammation, widespread decreases in Bolton-Hunter-labeled 125I-substance P binding occurred on both sides of the dorsal horn of spinal level L4 in comparison to the control group. However, by two days, widespread increases in Bolton-Hunterlabeled 125I-substance P binding occurred on both sides of the spinal cord at level L4 compared to the control group. The increase in radioligand binding was primarily due to a 10-fold increase in affinity of neurokinin-1 receptors for substance P. At later time-points of four and eight days, Bolton-Hunter-labeled 125I-substance P binding remained increased only in laminae I/II on the side of the spinal cord ipsilateral to inflammation. The changes in Bolton-Hunter-labeled 125I-substance P binding suggest that alterations in substance P synaptic transmission in the spinal cord may contribute to the increased excitability of spinal neurons that accompanies adjuvant-induced peripheral inflammation.