Ultrastructural characterization of axonal endings in the substantia gelatinosa which take up [3H]serotonin.

Serotonergic axonal endings in layers I and II of the dorsal horn of the medulla were identified by autoradiography. In adult cats, pretreated with a monoamine oxidase inhibitor, tritiated serotonin ([3H]5-HT) was topically applied onto the surface of the caudal medulla. Light autoradiographs from 1 micrometer sections demonstrated silver grains in both layers I and II. In EM autoradiographs, two categories of axonal endings were labeled by [3H]5-HT uptake: dome-shaped endings which form a single synapse and scalloped endings which form multiple synapses. Each category was further divided into several types based on morphological criteria. The [3H]5-HT-labeled endings synapse primarily on small caliber dendritic shafts and spines, with the dome-shaped endings forming both symmetrical and asymmetrical synapses and the scalloped endings forming only asymmetrical synapses. Dome-shaped endings were most common and two types were found in layers I and II while a third type was found only in layer II. Layer I contained a single type of scalloped ending while layer II contained three types of scalloped endings. In a series of experiments designed to provide another approach to identifying serotonergic endings, 5,6-dihydroxytryptamine, a serotonin neurotoxin, was either topically applied onto the caudal medulla or injected into the fourth ventricle. Following treatment with the neurotoxin, blackened degenerating dome-shaped and scalloped endings similar to those labeled in the [3H]5-HT uptake experiments were found in layers I and II. The presence of serotonergic endings in layer I suggests that some of these endings synapse on the dendrites of layer I projection neurons where they may inhibit the output of the projection neuron directly. Serotonergic endings in layer II may modulate the activity of layer II interneurons by synapsing directly on these interneurons. The interneurons in layer II may function by mediating the transfer of inputs from primary endings in these layers to layer I projection neurons.