This study examined the effect of cigarette smoking on the fluidity of the rat alveolar macrophage plasma membrane. Rats were subjected to 8 wk of an in vivo smoke exposure protocol, after which their alveolar macrophages were harvested. Fluidity was assessed by measuring steady-state anisotropy of isolated plasma membranes as well as of lipid vesicles made from total lipid extracts of these plasma membranes. The smoke-exposed animals showed a significant decrease in fluidity in both intact plasma membranes (p less than 0.0001) and in their lipid vesicle preparations (p less than 0.0001). To assess the time course of these changes, lipid vesicles were prepared from total cellular lipid extracts of macrophages from paired rats, control and smoke-exposed, at 1 through 4 wk after initiation of exposure. Significant decreases in fluidity were observed as early as 2 wk after smoking was begun (p less than 0.001). To assess the reversibility of these changes, paired rats were exposed for 8 wk, then withdrawn for 8, 12, and 18 wk, after which fluidity was evaluated in lipid vesicles prepared from total cellular lipids. Even after 18 wk of smoking cessation, significant decreases in fluidity persisted (p less than 0.01). We conclude that cigarette smoking causes a decrease in plasma membrane fluidity of rat alveolar macrophages. This is due at least in part to a change in the lipid portion of the membrane. These alterations occur after a very brief period of smoke exposure and persist long after cessation of smoking.