The aim of this study was to assess the effect of N-ethoxycarbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ)-induced alpha2-adrenoceptor inactivation on regulatory G proteins and the recovery of agonist and antagonist binding sites. EEDQ induced a rapid increase in the abundance of rat brain cortical Galphai1/2 proteins (30% at 6 h) which reached a maximum at 4 days (45%) and which then slowly returned (7-30 days) to control values. EEDQ did not alter significantly the levels of Galphai3 and Galphao proteins. By using the standard monoexponential model, the analysis of the recovery of alpha2-adrenoceptor density (6 h-30 days) with [3H]UK 14304 (bromoxidine) and [3H]RX 821002 (2-metoxy idazoxan) in the cerebral cortex did not reveal differences in receptor turnover parameters. However, the recovery of [3H]UK 14304 binding fitted best to a new biphasic recovery model, suggesting the existence of two distinct phases of recovery of agonist sites (r1 and r2 = 15.7 and 7.4 fmol mg protein(-1) day(-1); k1 and k2 = 0.51 and 0.25 day(-1); (t1/2)1 and (t1/2)2 = 1.4 and 2.7 days). In contrast, the recovery of [3H]RX 821002 antagonist sites did not fit to the biphasic model (r = 8.1, k = 0.14, t1/2 = 4.9). Because agonist binding requires coupling to G proteins, the present results suggest that the rapid over-expression of Galphai1/2 proteins induced by EEDQ is related to the biphasic recovery of [3H]UK 14304 binding. The possible implication of the faster recovery of alpha2-adrenoceptor function after EEDQ inactivation is discussed.