Direct binding studies on longitudinal ileal and atrial muscarinic receptors revealed that most of the ileal or atrial selective antagonists identified in functional studies did not differentiate between these muscarinic receptors in direct binding studies. Methoctramine, an atrial selective muscarinic receptor antagonist in functional studies was, however, able to partially discriminate between these two receptors in our binding studies. Furthermore the binding data obtained using this compound indicated that longitudinal ileal muscarinic receptors were heterogeneous. The predominant population of ileal muscarinic receptors displayed a similar pharmacology to the cardiac type M2 muscarinic receptor. The minor population of muscarinic receptors identified in binding studies displayed a similar pharmacology to the ileal muscarinic receptor identified in functional studies and were pharmacologically similar to the exocrine gland type M2 muscarinic receptor.