While retrograde regulation of neuronal development by target-derived factors in the autonomic nervous system is well established, the importance of anterograde influences on target development is unclear. Previous studies suggest that sympathetic innervation of sweat glands plays a critical role in the acquisition and maintenance of their secretory function. To define the signal(s) responsible, we disrupted muscarinic chollnergic transmission in developing and adult rats. Treatment of young rats with the nonselective antagonist, atropine, or an antagonist selective for the glandular muscarinic subtype, 4-DAMP, delayed the development of secretory responsiveness. Treatment of adult animals with atropine caused its loss. Further, following denervation, treatment with the muscarinic agonist, pilocarpine, largely preserved responsiveness while untreated animals lost function. Thus, acetylcholine, whose presence in sweat gland innervation is retrogradely specified by developmental interactions with the target tissue, In turn plays an important role in inducing and maintaining target tissue responsiveness through muscarinic receptor activation.