The occurrence of acetylcholine (ACh)-like activity in human placenta, a tissue without innervation, has been known for more than 60 years. However, the non-neuronal functions of ACh in human placenta are not clearly understood. The components of the cholinergic system-ACh, choline acetyltransferase, acetylcholinesterase, butyrylcholinesterase, muscarinic receptors, and nicotinic receptors--in human placenta have been demonstrated by unequivocal methods. Primate placentae store and release ACh by mechanisms similar to those of nervous tissue. However, there are many gaps in our knowledge, which include: (a) endogenous quaternary ammonium compounds other than ACh in human placental extracts; (b) the specificity of placental enzymes; (c) the subtypes and structures of placental muscarinic and nicotinic receptors; and (d) the significance of placental alpha-bungarotoxin binding proteins, ACh receptor stimulation-cellular signaling by second messengers, and activation of immediate early target genes (C-fos, C-jun) encoding transcription factors. Several hypothetical non-neuronal functions of ACh in placenta have been postulated based upon available experimental evidence. These include: (a) regulation of blood flow and fluid volume in placental vessels; (b) opening and closing of trophoblastic channels; (c) induction of contractile properties to myofibroblasts; (d) facilitation of amino acid transport necessary for fetal growth across placenta; (e) release of placental hormones; and (f) modulation of the formation of myometrial and placental prostaglandins in human parturition. All of these roles are reasonable, and some of these roles mav turn out to be linked to one another to influence or maintain placental function.