Role of L-type calcium channels on yohimbine-precipitated clonidine withdrawal in vivo and in vitro.

This study was designed to elucidate the possible participation of L-type calcium channels in the expression of clonidine-withdrawal precipitated by yohimbine in clonidine-dependent animals. Mice implanted for 5 days with osmotic minipumps containing the alpha 2-adrenoceptor agonist clonidine showed symptoms of a withdrawal syndrome (jerks, headshakes, defecations and weight loss) when yohimbine, an alpha 2-adrenoceptor antagonist, was injected. Similarly, isolated rat ilea incubated with clonidine in vitro showed a withdrawal contracture when yohimbine was added to the organ bath. The effects of L-type calcium channel blockers (verapamil and diltiazem) and the stimulant Bay K 8644 on these two different types of withdrawal responses were evaluated. A dose-dependent decrease in yohimbine-precipitated clonidine withdrawal in vivo was observed when verapamil (10-40 mg/kg, s.c. and 120 micrograms/mouse, i.c.v.) or diltiazem (5-20 mg/kg, s.c. and 160 micrograms/mouse, i.c.v.) were administered to mice dependent on clonidine. No effect was found after Bay K 8644 (0.5-5 mg/kg, s.c. and 1-5 micrograms/mouse) was injected under these conditions. In vitro, both verapamil (0.1-5 microM) and D-cis-diltiazem (1-50 microM) concentration-dependently reduced the height of the yohimbine-precipitated withdrawal contracture in rat ileum incubated with clonidine. Furthermore, the effect of diltiazem was stereospecific, as D-cis-diltiazem 10 microM markedly inhibited clonidine withdrawal, whereas the same concentration of L-cis-diltiazem had no effect. In contrast, the calcium channel stimulant Bay K 8644 (0.1-1 microM) increased the height of the ileum withdrawal contracture.(ABSTRACT TRUNCATED AT 250 WORDS)