We have devised a method for the parallel determination of choline acetyltransferase (CAT) and muscarinic cholinergic receptor (mCh-R) in the same brain tissue. The method for CAT activity determination is more rapid, simplified, stable, and economical than the usual Fonnum's method. With our method, age-associated changes in CAT activity and mCh-R levels were examined. Although CAT activity hardly changed with age except in a few areas, mCh-R binding of aged-rats was markedly reduced in all areas. These results suggest that the change in mCh-R represents an age-associated biochemical change in the brain and that determination of CAT activity is not sufficient for the study of age-associated changes in the brain cholinergic system.