The effects of locally administered physostigmine and di-isopropylphosphorofluoridate (DFP) were compared on the secretory response of sweat glands to intradermally injected acetylcholine and carbachol in healthy male volunteers (physostigmine: six subjects; DFP: one subject). The response to acetylcholine reached its peak within 10 s of injection and then rapidly declined, whereas the response to carbachol increased steadily reaching a peak between 5 and 7 min after injection. The response to acetylcholine was potentiated in the presence of both physostigmine and DFP, whilst the response to carbachol was not significantly affected by either of these drugs. The difference in the time-course of responses to acetylcholine and carbachol may be attributed to differences in the susceptibility of the two drugs to metabolism by acetylcholinesterase; carbachol, unlike acetylcholine, being virtually immune to metabolism by this enzyme. It is concluded that the response to carbachol is mediated by a direct stimulatory action on post-synaptic muscarinic receptors rather than by activation of pre-synaptic nicotinic receptors leading to the release of endogenous acetylcholine.