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1. The acetylcholine (ACh)-releasing system in the cerebral cortex of pentobarbital anaesthetized cats was investigated by examining the effect of graded afferent volleys in forelimb nerves on ACh release from the sensorimotor cortices contralateral and ipsilateral to the site of stimulation. 2. Cortical ACh release was determined by bio-assay of neostigmine-containing perfusates which had been in contact with the cortical surfaces for 5-10 min periods. 3. Afferent volleys, generated by stimuli that were effective in activating as many fibres of a fibre group as possible without stimulating fibres in the group with the next highest threshold for activation, were monitored from dorsal roots C7 or C8 before entering the spinal cord. 4. Stimulation of the deep (DR) and superficial (SR) radial nerves and the radial (R) nerve proximal to the junction of the DR and SR were effective in enhancing ACh release only when either group III or groups III and IV fibres were included in the afferent volley. 5. The rates of ACh release from the primary receiving area of the sensorimotor cortex contralateral to the site of stimulation did not differ from those from the same area of the ipsilateral sensorimotor cortex. 6. The pertinence of this data to the various hypotheses concerning the nature of the ACh-releasing pathways to the cerebral cortex is discussed.