Arteriolar smooth muscle Ca2+dynamics during blood flow control in hamster cheek pouch

Abstract

Intracellular calcium concentration ([Ca2+]i) governs the contractile status of arteriolar smooth muscle cells (SMC). Although studied in vitro, little is known of SMC [Ca2+]idynamics during the local control of blood flow. We tested the hypothesis that the rise and fall of SMC [Ca2+]iunderlies arteriolar constriction and dilation in vivo. Aparenchymal segments of second-order arterioles (diameter 35 ± 2 μm) were prepared in the superfused cheek pouch of anesthetized hamsters ( n = 18) and perifused with the ratiometric dye fura PE-3 (AM) to load SMC (1 μM, 20 min). Resting SMC [Ca2+]iwas 406 ± 37 nM. Elevating superfusate O2from 0 to 21% produced constriction (11 ± 2 μm) that was unaffected by dye loading; [Ca2+]iincreased by 108 ± 53 nM ( n = 6, P < 0.05). Cycling of [Ca2+]iduring vasomotion (amplitude, 150 ± 53 nM; n = 4) preceded corresponding diameter changes (7 ± 1 μm) by ∼2 s. Microiontophoresis (1 μm pipette tip; 1 μA, 1 s) of phenylephrine (PE) transiently increased [Ca2+]iby 479 ± 64 nM ( n = 8, P < 0.05) with constriction (26 ± 3 μm). Flushing blood from the lumen with saline increased fluorescence at 510 nm by ∼45% during excitation at both 340 and 380 nm with no difference in resting [Ca2+]i, diameter or respective responses to PE ( n = 7). Acetylcholine microiontophoresis (1 μA, 1 s) transiently reduced resting SMC [Ca2+]iby 131 ± 21 nM ( n = 6, P < 0.05) with vasodilation (17 ± 1 μm). Superfusion of sodium nitroprusside (10 μM) transiently reduced SMC [Ca2+]iby 124 ± 18 nM ( n = 6, P < 0.05), whereas dilation (23 ± 5 μm) was sustained. Resolution of arteriolar SMC [Ca2+]iin vivo discriminates key signaling events that govern the local control of tissue blood flow.