Meeting Report for: The 8th International Symposium on Cholinergic Mechanisms

Based on the sequence of the five cloned muscarinic receptor subtypes (m1-m5), subtype selective antibody and cDNA probes have been prepared. Use of these probes has demonstrated that each of the five subtypes has a markedly distinct distribution within the brain and among peripheral tissues. The distributions of these subtypes and their potential physiological roles are discussed. By use of molecular genetic manipulation of cloned muscarinic receptor cDNAs, the regions of muscarinic receptors that specify G-protein coupling and ligand binding have been defined in several recent studies. Overall, these studies have shown that amino acids within the third cytoplasmic loop of the receptors define their selectivities for different G-proteins and that multiple discontinuous epitopes contribute to their selectivities for different ligands. The residues that contribute to ligand binding and G-protein coupling are described, as well as the implied structures of these functional domains.

It could be argued that clinical experience with cholinergic drugs in the therapy of AD has not yet shown relevant symptomatic improvements. The main reasons for this might be attributed to peripheral cholinergic effects and the liver toxicity of some of these drugs, which limit their use and prevent confirmation of the cholinergic hypothesis (Gray et al., 1989). The main disadvantages of the cholinesterase inhibitors used in clinical trials are the short duration of action in the case of physostigmine and the potential for liver toxicity seen with the aminoacridine derivatives. The results presented with SDZ ENA 713 indicate that the disadvantages of AChE inhibitors might be overcome by improving CNS selectivity and thereby decreasing the peripheral cholinergic effects and toxicity. Clinico-pharmacological studies with SDZ ENA 713 have been performed in healthy volunteers; while central activity was clearly demonstrated in an EEG-sleep study (Holsboer et al., 1992), no prohibitive peripheral side effects were seen, confirming in humans the results obtained in experimental animals (Enz et al., 1991). A multicentre clinical investigation in AD patients has been performed in Europe and is currently being evaluated.

As reviewed here and elsewhere in this symposium, acetylcholine, in conjunction with other neurotransmitter systems, plays a very important role in the regulation of circadian and sleep-wake states. To briefly recapitulate, several current basic concepts about the regulation of sleep-wake states include: (a) REM sleep, or at least its phasic events (eye movements and PGO spikes), are promoted by cholinergic neurons originating within the peribrachial regions [LDT/PPT] (Mitani et al., 1988; Shiromani et al., 1988; Datta et al., 1991; Shouse and Siegel, 1992); (b) REM sleep may be inhibited by noradrenergic and serotonergic neurons in the locus coeruleus and dorsal raphe, respectively (Siegel, 1989; Steriade and McCarley, 1990; Jones, 1991); (c) stages 3 and 4 (Delta) sleep are inhibited by cholinergic terminals from basal forebrain to cortex (Buzsaki et al., 1988) and from LDT/PPT to thalamus (Steriade and McCarley, 1990; Steriade et al., 1991); (d) Delta sleep is modulated by complex serotonergic mechanisms; for example, it is increased by pharmacological antagonists of 5HT2 receptors (Declerck et al., 1987; Dugovic et al., 1989; Benson et al., 1991), although the mechanism and neuroanatomical site at which this effect occurs is unknown. Given the importance of mACHR mediation of components of REM sleep, it is unfortunate that so little is known about the distribution of the various subtypes of mACHRs in brainstem areas which regulate REM sleep. mACHR subtypes have been identified by molecular, biological and pharmacological methods.

Since the demonstration some 50 years ago of the presence and synthesis of acetylcholine (ACh) in specific neuronal systems within the brain, a wealth of information concerning the organization and functional importance of central cholinergic neurons has emerged through immunohistochemical, neuroanatomical, pharmacological, biochemical and neurophysiological studies. Many of the original theses have proven valid concerning the key structural and functional position of cholinergic neurons within the central reticular core of the brain, where the basic sleep-waking cycle is determined. The two major cholinergic cell groups of this core, one within the pontomesencephalic tegmentum that projects rostrally into the non-specific thalamo-cortical relay system and the other within the basal forebrain that receives input from the brainstem reticular formation and projects in turn as the ventral, extrathalamic relay upon the cerebral cortex, are critically involved in processes of cerebral activation that accompany the states of wakefulness and paradoxical sleep. By interaction with other cell groups, including monoaminergic and GABAergic neurons, and by differential modes of firing, the cholinergic neurons may furthermore shape the responsiveness and activity of the reticular core and thalamo-cortical systems across the sleep-waking cycle.