Synaptic contacts between serotonergic and cholinergic neurons in the rat dorsal raphe nucleus and laterodorsal tegmental nucleus

Abstract

We examined synaptic connectivity between cholinergic and serotonergic neurons in the dorsal raphe nucleus and the laterodorsal tegmental nucleus of the rat. To this purpose we employed two variations (the combination of pre-embedding immunogold–silver intensification with avidin–biotin-peroxidase complex technique and the combination of avidin–biotin–peroxidase/3,3′-diaminobenzidine/silver–gold intensification with avidin–biotin–peroxidase/3,3′-diaminobenzidine reaction) of a double pre-embedding immunoelectron procedure, using primary antibodies against vesicular acetylcholine transporter and serotonin. At the light-microscopic level, serotonin-like immunoreactive neurons in the dorsal raphe nucleus appeared as reddish black and vesicular acetylcholine transporter-like immunoreactive axon terminals were brown colored using a combination of pre-embedding immunogold-silver technique and avidin–biotin–peroxidase complex technique. Serotonin-like immunoreactive fibers projected to the laterodorsal tegmental nucleus. At the electron microscopy level, with both methods we observed in the dorsal raphe nucleus vesicular acetylcholine transporter-immunopositive axon terminals in synaptic contact with serotonin-like immunoreactive dendrites and, to a lesser degree, with serotonin-like immunoreactive cell bodies. These synapses usually were of the symmetrical type. Occasionally we noted, next to vesicular acetylcholine transporter-immunopositive axon terminals, also immunonegative terminals synapsing with the serotonin-like immunoreactive dendrites. In the laterodorsal tegmental nucleus we found serotonin-like immunoreactive axon terminals and immunonegative terminals forming synapses with vesicular acetylcholine transporter-immunoreactive dendrites. Most synapses formed by the serotonin-like immunopositive terminals were of the asymmetrical type. Our results suggest that serotonergic neurons in the dorsal raphe nucleus and cholinergic neurons in the laterodorsal tegmental nucleus may reciprocally influence each other by means of synaptic connectivity. Such connectivity may serve to regulate pain sensation, or be involved in the regulation of the sleeping–waking cycle.