Quantitative light microscopic autoradiographic localization of cholinergic muscarinic receptors in the human brain: brainstem

Abstract

We have investigated the localization of muscarinic cholinergic receptors in the brainstem of eight patients free of neurological disease following quantitative autoradiography of microtome sections of postmortem tissue labeled in vitro with N-[3H]methyl scopolamine as a ligand. Receptor densities were quantified by microdensitometry with the aid of a computer assisted image analysis system. Our results reveal a heterogeneous distribution of receptor sites. High concentrations of muscarinic cholinergic receptor sites were associated with many nuclei and areas of the brainstem including the nucleus facialis (VII), hypoglossus (XII), ambiguus, the motor trigeminal nucleus (V), the nucleus solitarius, the nucleus of the lateral lemniscus, the superior and inferior colliculi, the sensory trigeminal nucleus (substantia gelatinosa), the pontine nuclei, the parabrachial nuclei, some tegmental nuclei and the periaqueductal gray matter. Very high concentrations of N-[3H]methyl scopolamine binding sites were also localized in the ventral tegmental area, the nucleus paranigralis and the nucleus ovalis. Receptor densities varied between individual brains although the relative distribution of the densities in the different nuclei was the same for all of the brains examined. Most of the brainstem nuclei containing muscarinic cholinergic receptors were enriched in high affinity agonist binding sites as shown by characteristic displacement of the ligand with carbachol. Exceptions were the substantia nigra, the nucleus olivaris inferior and the substantia gelatinosa of the fifth nerve. Receptor density values and pharmacological characteristics obtained in the cortex and basal ganglia in our cases are in good agreement with previously reported values in humans, using conventional biochemical methods. This indicates that procedures used in the autoradiographic technique are not detrimental to the pharmacological characteristics and densities of muscarinic cholinergic receptors. Our results thus clearly show the feasibility of using these techniques for the localization and quantification of muscarinic cholinergic receptors in human brain postmortem material. Furthermore, our findings indicate the potential involvement of the muscarinic cholinergic effect of acetylcholine in the normal function of many brainstem centers, including motor and sensory nuclei, visual and auditory relay nuclei and cardiovascular and respiratory-related nuclei.