Fluorescent probes were used to compare the physical properties of membranes from mice selected for sensitivity (LS) and insensitivity (SS) to the hypnotic action of ethanol. Brain synaptic plasma membranes (SPM) from LS mice were more sensitive to the disordering action of ethanol than those from LS mice when probes were located near the membrane surface. However, the membrane core of membranes from the two lines was equally sensitive to ethanol. The genetic differences in ethanol sensitivity of the membrane surface were eliminated when fluorescence measurements were carried out in the presence of 2-3 mM CaCl2. Consistent with behavioral data, differential genetic sensitivity to the disordering action was not obtained with longer chain alcohols. The genetic difference in ethanol sensitivity was not detected with erythrocyte membranes or lipids extracted from SPM. These results indicate that there is a structural difference in the surface of brain membranes of LS and SS mice than may influence their sensitivity to ethanol.