Acetylcholine was the first neurotransmitter identified and ATP is the hitherto final compound added to the list of small molecule neurotransmitters. Despite the wealth of evidence assigning a signaling role to extracellular ATP and other nucleotides in neural and non-neural tissues, the significance of this signaling pathway was accepted very reluctantly. In view of this, this short commentary contrasts the principal molecular and functional components of the cholinergic signaling pathway with those of ATP and other nucleotides. It highlights pathways of their discovery and analyses tissue distribution, synthesis, uptake, vesicular storage, receptors, release, extracellular hydrolysis as well as pathophysiological significance. There are differences but also striking similarities. Comparable to ACh, ATP is taken up and stored in synaptic vesicles, released in a Ca(2+)-dependent manner, acts on nearby ligand-gated or metabotropic receptors and is hydrolyzed extracellularly. ATP and acetylcholine are also costored and coreleased. In addition, ATP is coreleased from biogenic amine storing nerve terminals as well as from at least subpopulations of glutamatergic and GABAergic terminals. Both ACh and ATP fulfill the criteria postulated for neurotransmitters. More recent evidence reveals that the two messengers are not confined to neural functions, exerting a considerable variety of non-neural functions in non-innervated tissues. While it has long been known that a substantial number of pathologies originate from malfunctions of the cholinergic system there is now ample evidence that numerous pathological conditions have a purinergic component.