Steady-state pHi, buffering power, and effect of CO2 in a smooth muscle-like cell line

Abstract

Intracellular pH (pHi) was studied in the smooth muscle-like cell line, BC3H-1, using the pH-sensitive fluorescent dye 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein (BCECF). The initial pHi measured in 20 mM Na N-2-hydroxyethylpiperazine-N'-2 ethanesulfonic acid-buffered medium [NHB; external pH (pHo) 7.4, 37 degrees C] was 6.89 +/- 0.01 (n = 178). pHi was affected by changes in external pHo, pHi changing by approximately 70% of the change in pHo. The intrinsic buffering power (beta int) of these cells, measured either with NH4Cl or Na propionate pulses, is low for muscle cells, averaging approximately 10 mM/pH unit. Steady-state pHi of BC3H-1 cells in NHB acidified reversibly on exposure to 0.5 mM 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS; 1.4 +/- 0.3 x 10(-4) pH/s), 1 mM amiloride (2.0 +/- 0.7 x 10(-4) pH/s), or Na-free solution (8.3 +/- 2.4 x 10(-4) pH/s) and alkalinized upon exposure to Cl-free solutions (9.7 +/- 2.2 x 10(-4) pH/s). Exposure of BC3H-1 cells to CO2-HCO3-buffered solutions resulted in a transient acidification followed by an alkalinization of 0.3-0.4 pH unit to a new steady-state pHi of 7.27 +/- 0.01 (n = 65). This new steady-state pHi acidified very slowly upon exposure to 1 mM amiloride (0.3 +/- 0.1 x 10(-4) pH/s), acidified more rapidly upon exposure to 0.5 mM DIDS (5.9 +/- 0.6 x 10(-4) pH/s) or Na-free solutions (9.8 +/- 1.0 x 10(-4) pH/s), and alkalinized on exposure to Cl-free solutions (24.5 +/- 1.3 x 10(-4) pH/s).(ABSTRACT TRUNCATED AT 250 WORDS)