Intact brain cell aggregates were dissociated from adult rat brains, by a simple sieving technique, and were used to study the binding characteristics of [3H]N-methylscopolamine to muscarinic acetylcholine receptors. The magnitude of binding of this ligand was related linearly to the amount of cell protein in the binding assay, with a high ratio of total to nonspecific binding. In addition, specific binding showed saturability and high affinity. Muscarinic receptor antagonists displaced specific [3H]N-methylscopolamine binding according to the law of mass-action, while it was possible to resolve displacement curves using receptor agonists into high- and low-affinity components. The results are discussed in terms of the usefulness of dissociated intact rat brain cells in studying muscarinic acetylcholine receptors in the central nervous system.