The density of substance P (SP) and serotonin (5HT) immunoreactivity in laminae I and II of rat spinal cord changes following dorsal rhizotomy in a manner consistent with sprouting by intrinsic SP and descending 5HT systems (Wang et al. J. Comp. Neurol. 304: 555, 1991). In this study we used quantitative EM-immunocytochemistry to examine whether the increase in substance P and serotonin immunoreactivity seen at the light microscopic level was related to increased numbers of SP- and 5HT-containing terminals in lamina II. Dorsal roots were sectioned and their ganglia removed from L1 to S2 unilaterally in 18 rats. After 3 (acute), 10 (subacute), or 60 (chronic) days, rats were perfused, and the L5 segments of the spinal cord were removed and prepared for electron microscopy. Lumbosacral deafferentation completely and permanently eliminated complex terminals in lamina II at L5 but the number of simple terminals increased by 46% compared to the control side. This result was similar to that shown previously in cat (Murray and Goldberger, J. Neurosci. 6: 3205, 1986) and suggested that intact intrinsic systems sprouted to form new terminals to compensate for the terminals lost by deafferentation. Quantitative electron microscopic immunocytochemistry demonstrated that the number of terminals containing SP in deafferented lamina II decreased by 58% at 3 days post-operatively and then increased by 10 days and recovered to normal levels by 60 days. The loss of SP terminals in the acute group is due to the loss of SP-containing dorsal root afferents, while the recovery in the chronic group suggests replacement of lost terminals by intrinsic SP systems. These results therefore indicate that SP-containing terminals show homotypic sprouting in response to complete dorsal root deafferentation. The number of 5HT-containing terminals in lamina II of spinal cord increased by 56% on the deafferented side in the chronic group. The increase in 5HT-containing terminals indicates that descending 5HT systems undergo heterotypic sprouting in response to dorsal rhizotomy.