Endothelial cells (EC) cultured on polymerized silicone deform the underlying substrate, producing microscopically visible wrinkles. This has been interpreted as cellular contraction, and we have previously concluded that EC normally maintain an active contractile tone. Since in ischemic tissues capillaries become "paralyzed" and lose their tone, we decided to examine the effects of glucose and/or oxygen deprivation on EC contractility. Contracting cultures with wrinkled silicone substrates were exposed to complete anoxia with or without exogenous glucose and followed by time-lapse photography. Under either glucose-or oxygen-free conditions, contraction was maintained for up to 4 days. If, however, both oxygen and glucose were removed, cellular contraction was reversed. After a period of 2 to 4 hours substrate wrinkles gradually disappeared, until by 3 to 7 hours, few to no wrinkles remained. Furthermore, within 10 minutes of restoration to normal oxygen (but not glucose) levels, substrate wrinkling reappeared. F-actin microfilament patterns and cell number per unit area were also altered by glucose and oxygen deprivation. Similar results were obtained using large or small vessel EC. We conclude that in the absence of glucose and oxygen EC lose their contractile tone, and that tone can be re-established upon re-exposure to oxygen. These findings should have implications for the pathogenesis of capillary paralysis in ischemia.