Effects of extravascular acidification and extravascular alkalinization on constriction and depolarization in rat cerebral arterioles in vitro

Abstract

✓ The relationship between cell membrane potential, vessel diameter, and pH in small cerebral arterioles is not completely understood. This study involved direct, simultaneous measurement of cell membrane potential and vessel diameter at various extracellular pH levels. Arterioles ranging from 44 to 91 µm in diameter were isolated, transferred to a temperature-controlled microscope chamber, which was used as an organ bath, and observed through an inverted videomicroscope. Two vessel cannulation procedures were used: a single-sided cannulation with the other side occluded, and a double-sided and perfused cannulation. After cannulation, the vessels were pressurized to 60 mm Hg intraluminally and the bath temperature was raised to 37°C. Cell membrane potentials of vessel wall cells were obtained after the bath temperature reached 37°C with the vessels partly constricted and again after spontaneous tone (constriction) of the healthy vessels had developed. The effect of extraluminal pH on cell membrane potentials was studied by changing the bath pH from 7.3 to either 7.65 or 6.8 in the single-sided cannulation. The average cell membrane potential for vessels at 37°C, with 60 mm Hg of intraluminal pressure and pH 7.3, was −37.5 mV. The cell membrane potential depolarized to −30.9 mV at pH 7.65 and hyperpolarized to −58.4 mV at pH 6.8, with a slope of 25.8 mV/pH unit. The effect of depolarizing extracellular potassium ions on the cell membrane potential was examined by perfusing two vessels with modified Ringer's solution containing 70 mM KCl. This perfusion method decreased the vessel diameter by 48% and depolarized the observed cell membrane potential from −41.9 to −19.8 mV, with a slope of −0.42 mV per percentage diameter change. These data provide the first measurements of membrane potentials of isolated penetrating arteriole wall cells in vitro. The results indicate that the cell membrane potential relates linearly to the vessel diameter. This new technique opens the possibility for studying vessel response to stimuli under controlled conditions and regulatory mechanisms such as the propagation of vasomotor responses.