Role of Ca2+- and swelling-activated Cl- channels in alpha(1)-adrenoceptor-mediated tone in pressurized rabbit mesenteric arterioles

Background: Ca2+-activated (I-Cl(Ca)) and swelling-induced (I-Cl(swell)) Cl - channels have, respectively, been postulated to participate in the membra ne depolarization and contraction mediated by activation of alpha(1)-adreno ceptors and vascular wall distension during pressurization. Their respectiv e function in generating active force in pressurized arterioles during alph a(1)-adrenoceptor stimulation remains unsettled. Objectives: Experimental p rotocols were designed to: (1) assess the relative contribution of I-Cl(Ca) to the pressure-dependence of lumen diameter of mesenteric arterioles at d ifferent states of activation of the alpha(1)-adrenoceptor, and (2) investi gate the potential role of I-Cl(Ca) in spontaneous and agonist-mediated myo genic reactivity. Methods: Segments of endothelium-denuded rabbit mesenteri c arterioles with a lumen diameter of similar to 70 mu m were cannulated at both ends and studied under isobaric conditions at 36 degrees C. Steady-st ate lumen diameter at each pressure step investigated (0-100 mmHg, in 20-mm Hg increments) was measured by a video-microscopy edge-detection technique. Results: Under control conditions, 23% of the arterioles developed nifedip ine-sensitive spontaneous myogenic tone. In the presence of 1 mM tetraethyl ammonium chloride (TEA) to inhibit Ca2+-dependent K+ channels, the alpha(1) -agonist phenylephrine (PE) contracted the vessels in a concentration-depen dent manner (0.1-10 mu M) and potentiated myogenic reactivity. The contract ion mediated by 1 mu M PE/TEA was abolished by 1 mu M nifedipine, indicatin g that Ca2+ entry through voltage-gated Ca2+ channels was a necessary step in the cascade leading to contraction. Niflumic acid (NfA, 100 mu M), a rel atively selective inhibitor of I-Cl(Ca) had no effect on myogenic tone but reversed the PE-induced contraction, varying with the concentration of PE a nd transmural pressure. For PE concentrations between 0.1 and 1 mu M, but n ot for 10 mu M PE, the relaxing efficacy of NfA decreased as applied pressu re was raised from 0 to 100 mmHg. At all pressure steps, the NfA-induced re laxation was inversely related to the concentration of PE. DIDS (200 mu M), another Cl- channel blocker, inhibited spontaneous myogenic tone, and part ially suppressed a component of contraction at elevated transmural pressure s in arterioles incubated in 1 mu M PE/1 mM TEA/100 mu M NfA. Conclusions: Our data indicate that under low to moderate stimulation of the alpha(1)-ad renoceptor signaling pathway, I-Cl(Ca) channels play an important role in t he sustained contraction produced. Their declining contribution to contract ion with increasing transmural pressure may be explained, at least in part, by a progressive enhancement of stretch-induced ionic conductances, possib ly volume-sensitive Cl- channels. (C) 2000 Elsevier Science B.V. All rights reserved.