Choline plays a central role in many physiological pathways, including neurotransmitter synthesis (acetylcholine), cell-membrane signaling (phospholipids), lipid transport (lipoproteins), and methyl-group metabolism (homocysteine reduction). Endurance exercise might stress several of these pathways, increasing the demand for choline as a metabolic substrate. This review examines the current literature linking endurance exercise and choline demand in the human body. Also reviewed are the mechanisms by which exercise might affect blood choline levels, and the links between methyl metabolism and the availability of free choline are highlighted. Finally, the ability of oral choline supplements to augment endurance performance is assessed. Most individuals consume adequate amounts of choline, although there is evidence that current recommendations might be insufficient for some adult men. Only strenuous and prolonged physical activity appears sufficient to significantly decrease circulating choline stores. Moreover, oral choline supplementation might only increase endurance performance in activities that reduce circulating choline levels below normal.