M1 Binding Sites Research Articles

Cholinergic binding in the hippocampus of the aging male rat

1. 1. M1 muscarinic ( 3-pirenzepine) and 3H- l-nicotine binding were measured in the hippocampus of male Wistar rats aged 3–4, 10–11 and 24–25 months. 2. 2. The maximal number of M1 binding sites did not differ between age groups. 3. 3. The dissociation constant of M1 binding was higher in old rats than in young rats. 4. 4. The binding of 3H- l-nicotine did not differ between age groups. 5. 5. The number of postsynaptic muscarinic receptors may be preserved, but the conformation of these receptors in the rat hippocampus may be altered during aging.

Muscarinic cholinergic receptor subtypes in normal human brain and Alzheimer's presenile dementia

Brain muscarinic M1 and M2 binding sites, as defined by their affinity for pirenzepine, were studied in 24 regions of 3 normal control brains. For each region, the binding of [ 3H]QNB, a non-subtype selective antagonist, was saturable with a mean K D of 0.117 ± 0.066 nM. Computer-assisted analysis of the pirenzepine competition binding curves yielded the amount of high affinity (M1) and low affinity sites (M2) as well as their respective K i value for this ligand. The neocortex contained a mixed population of 67% M1 and 33% M2 sites, without locoregional heterogeneity. The muscarinic receptors of the caudate nucleus, putamen, pallidum, hippocampus and nucleus amygdalis were predominantly of the M1 type as well. The cerebellum and the brain-stem are examples of regions containing over 90% M2 sites. White matter structures like the centrum ovale appeared to contain low concentrations of [ 3H]QNB binding sites, predominantly of the M1 subtype. The data for the frontal cortex were compared with data obtained in 3 patients who died from Alzheimer's disease with very early onset: the muscarinic receptor concentrations were somewhat lower but their M1 M2 ratio remained unchanged. GTP caused an appreciable rightward shift and steepening of the carbachol competition binding curve in M2 predominant regions such as the pons, whereas only a slight shift was observed in the cortex. In the presence of GTP, the alkylating reagent N-ethylmaleimide caused a 35-fold increase of the affinity for carbachol in M2 predominant regions. In contrast, regions where the receptors are predominantly of the M1 type, N-ethylmaleimide caused only a 5-fold increase in agonist affinity. These findings confirm our previously formulated hypothesis that the ability of N-ethylmaleimide to modulate the agonist affinity is an additional criterion for the characterization of M1 and M2-type receptors.

Facilitation of amphetamine-induced rotation by muscarinic antagonists is correlated with M 2 receptor affinity

This study examined the relationship between the affinity of cholinergic drugs for muscarinic receptor subtypes and their potency in potentiating or inhibiting amphetamine-induced rotation. The ascending nigrostriatal dopaminergic pathway was unilaterally lesioned in male Wistar rats using 6-hydroxydopamine. In these rats, ipsiversive rotation induced by amphetamine sulphate (1 mg/kg, s.c.) was dose-dependently inhibited by the cholinergic agonists oxotremorine, RS86 and pilocarpine and by the acetylcholinesterase inhibitor physostigmine. In contrast the cholinergic antagonists scopolamine, secoverine and dicyclomine facilitated amphetamine-induced rotation. Agonist and antagonist potencies were then compared with M1 and M2 binding site affinities estimated by displacing [3H]pirenzepine from forebrain and [3H]QNB from brainstem homogenates. The data suggest a relationship between antagonist potency and M2 binding site affinity.

Muscarinic activity of McN‐A‐343 and its value in muscarinic receptor classification

The affinity and potency of McN-A-343 (4-(m-chlorophenyl-carbamoyloxy) -2-butynyltrimethylammonium chloride) has been assessed at a range of M1 and M2 muscarinic receptors. McN-A-343 was shown to act as a full agonist at M2 receptors present in the guinea-pig isolated taenia caeci (-log EC50 = 5.14). McN-A-343 exhibited no agonist action in the guinea-pig ileum, atria, bladder or trachea. McN-A-343 was not selective in terms of affinity since its dissociation constants at M1 and M2 binding sites in the rat cerebral cortex and myocardium respectively, were very similar (cortical pPKi = 5.05; myocardial pKi = 5.22). The selectivity previously reported for the compound may be due to differences in intrinsic efficacy and/or tissue receptor reserve. Based on differential antagonist affinities, the muscarinic receptor profile of the taenia caeci, trachea and bladder was similar to that observed in the ileum, but dissimiliar to that observed in the atria.