Frequency receptive fields (RFs) were determined before and after pairing iontophoretic administration of acetylcholine (ACh) with a repeated single-frequency stimulus in the auditory cortex of barbiturate-anesthetized cats. In 58% of the cells, the paired ACh + tone treatment produced subsequent alterations of frequency RFs. In half of these cases, the RF modifications were highly specific to the frequency that had been paired with ACh. Atropine antagonized the frequency-specific effects of ACh, suggesting that they were mediated via muscarinic cholinergic receptors.