Cytosolic calcium concentration in resting and stimulated endothelium of excised intact rat aorta

1. Optical fibres were used to excite and record fluorescence from the lumenal face of rat aorta or tail artery loaded with fura-2. 2. Acetylcholine (ACh) evoked an endothelium-dependent rise in the fura-2 340/380 nm excitation ratio in both vessels. High [K+] or phenylephrine evoked an endothelium-independent rise in ratio in tail artery but failed to increase the ratio in aorta. These observations indicate that fura-2 fluorescence and therefore cytosolic calcium concentration ([Ca2(+)](i)) may be selectively recorded from the endothelium of intact rat aorta. 3. In aortic endothelium, resting [Ca2+](i) was 95 +/- 8 nM (n = 44). ACh evoked a monophasic rise in [Ca2+](i) which was temporally coincident with a membrane hyperpolarization. 4. BTP in most (22/35) preparations evoked a rise in [Ca2+](i) which declined towards resting and was followed by a secondary rise. The biphasic [Ca2+](i) responses were accompanied by biphasic electrical responses of initial hyperpolarization followed by depolarization above the resting potential and subsequent restoration towards rest. In the presence of high [K+] or the K+ ionophore valinomycin, ATP did not evoke changes in membrane potential and only monophasic rises in [Ca2+](i) were observed. In some (7/35) preparations, ATP evoked oscillations in [Ca2+](i), with membrane potential oscillating in antiphase. 5. These data suggest interplay between [Ca2+](i) and membrane potential in the generation of agonist-evoked responses in native endothelium in situ. The observed oscillations in [Ca2+](i) imply spatio-temporal synchronization of Ca2+ signalling in large groups of endothelial cells in intact rat aorta.