Cannabinoid CB1 receptor and endothelium‐dependent hyperpolarization in guinea‐pig carotid, rat mesenteric and porcine coronary arteries
Authors: Chataigneau T , Félétou M , Thollon C , Villeneuve N , Vilaine J‐ P. , Duhault J , Vanhoutte P M. 
Published: January 2005

The purpose of these experiments was to determine whether or not the endothelium‐dependent hyperpolarizations of the vascular smooth muscle cells (observed in the presence of inhibitors of nitric oxide synthase and cyclo‐oxygenase) can be attributed to the production of an endogenous cannabinoid.

Membrane potential was recorded in the guinea‐pig carotid, rat mesenteric and porcine coronary arteries by intracellular microelectrodes.

In the rat mesenteric artery, the cannabinoid receptor antagonist, SR 141716 (1 μM), did not modify either the resting membrane potential of smooth muscle cells or the endothelium‐dependent hyperpolarization induced by acetylcholine (1 μM) (17.3±1.8 mV, n=4 and 17.8±2.6 mV, n=4, in control and presence of SR 141716, respectively). Anandamide (30 μM) induced a hyperpolarization of the smooth muscle cells (12.6±1.4 mV, n=13 and 2.0±3.0 mV, n=6 in vessels with and without endothelium, respectively) which could not be repeated in the same tissue, whereas acetylcholine was still able to hyperpolarize the preparation. The hyperpolarization induced by anandamide was not significantly influenced by SR 141716 (1 μM). HU‐210 (30 μM), a synthetic CB1 receptor agonist, and palmitoylethanolamide (30 μM), a CB2 receptor agonist, did not influence the membrane potential of the vascular smooth muscle cells.

In the rat mesenteric artery, the endothelium‐dependent hyperpolarization induced by acetylcholine (1 μM) (19.0±1.7 mV, n=6) was not altered by glibenclamide (1 μM; 17.7±2.3 mV, n=3). However, the combination of charybdotoxin (0.1 μM) plus apamin (0.5 μM) abolished the acetylcholine‐induced hyperpolarization and under these conditions, acetylcholine evoked a depolarization (7.7±2.7 mV, n=3). The hyperpolarization induced by anandamide (30 μM) (12.6±1.4 mV, n=13) was significantly inhibited by glibenclamide (4.0±0.4 mV, n=4) but not significantly affected by the combination of charybdotoxin plus apamin (17.3±2.3 mV, n=4).

In the guinea‐pig carotid artery, acetylcholine (1 μM) evoked endothelium‐dependent hyperpolarization (18.8±0.7 mV, n=15). SR 141716 (10 nM to 10 μM), caused a direct, concentration‐dependent hyperpolarization (up to 10 mV at 10 μM) and a significant inhibition of the acetylcholine‐induced hyperpolarization. Anandamide (0.1 to 3 μM) did not influence the membrane potential. At a concentration of 30 μM, the cannabinoid agonist induced a non‐reproducible hyperpolarization (5.6±1.3 mV, n=10) with a slow onset. SR 141716 (1 μM) did not affect the hyperpolarization induced by 30 μM anandamide (5.3±1.5 mV, n=3).

In the porcine coronary artery, anandamide up to 30 μM did not hyperpolarize or relax the smooth muscle cells. The endothelium‐dependent hyperpolarization and relaxation induced by bradykinin were not influenced by SR 141716 (1 μM).

These results indicate that the endothelium‐dependent hyperpolarizations, observed in the guinea‐pig carotid, rat mesenteric and porcine coronary arteries, are not related to the activation of cannabinoid CB1 receptors.

British Journal of Pharmacology (1998) 123, 968–974; doi:10.1038/sj.bjp.0701690