Autoregulation of cholinergic neurons in the laterodorsal tegmental (LDT) and pedunculopontine (PPT) nuclei has been implicated in many functions, most importantly in drug reinforcement and in the pathophysiology of schizophrenia. This autoregulation is attributed to the release of acetylcholine, but neither the storage or release sites are known. To determine these sites, we used electron microscopy for the immunocytochemical localization of antipeptide antiserum raised against the vesicular acetylcholine transporter (VAchT) that is responsible for the uptake of acetylcholine into storage vesicles. The cellular and subcellular distribution of VAchT was remarkably similar in the two regions by by using each of two methods, immunogold and immunoperoxidase. In both PPT and LDT nuclei, VAchT labeling was seen mainly on membranous organelles including the trans-Golgi network in many somata. VAchT-immunoreactive tubulovesicles resembling saccules of smooth endoplasmic reticulum were often seen near the plasma membrane in dendrites. The VAchT-containing dendrites comprised almost 50% of the labeled profiles (1027/2129) in PPT and LDT nuclei. The remaining VAchT-immunoreactive profiles were primarily small unmyelinated axons and axon terminals. In axon terminals, VAchT was densely localized to membranes of small synaptic vesicles. The VAchT-immunoreactive axon terminals formed either symmetric or asymmetric synapses. The postsynaptic targets of these axon terminals included dendrites that were with (36/110) or without (74/110) VAchT immunoreactivity. Our results suggest that dendrites, as well as axon terminals, have the potential for storage and release of acetylcholine in the LDT and PPT nuclei. The released acetylcholine is likely to play a major role in autoregulation of mesopontine cholinergic neurons.
Copyright 2000 Wiley-Liss, Inc.