Vaughan-Jones R D
J Physiol. 1986 Oct;379:377-406. doi: 10.1113/jphysiol.1986.sp016259.
Intracellular Cl activity (aiCl), and intracellular pH (pHi) were measured in isolated sheep cardiac Purkinje fibres using a liquid ion exchanger Cl-selective micro-electrode and a glass recessed-tip, pH-selective micro-electrode. Removal of external Cl (glucuronate substituted) produced a fall in aiCl from about 20 to about 4 mmol/l: the residual level is probably caused by intracellular interference on the Cl-sensitive electrode. Re-exposure of the fibre to increased levels of external Cl produced, in the steady state, increased levels of aiCl. The dependence of steady-state aiCl upon external Cl activity, aoCl, was roughly hyperbolic with 50% recovery occurring at an aoCl of about 9.5 mmol/l. At all levels of external Cl tested, Cl was accumulated to a level much higher than that predicted for passive electrochemical equilibrium. Exposure of a Cl-depleted fibre to various levels of external Cl produced an exponential rise with time in aiCl. The initial rate-of-rise in aiCl was estimated to be a saturating function of aoCl, with a half-maximal effect occurring at an aoCl of about 33 mmol/l. The rate-of-rise was about 10-fold greater than that predicted from constant-field theory using published values for PCl, the Cl permeability coefficient. Steady-state aiCl was essentially insensitive to changes in external HCO3 concentration, [HCO3]o, if these changes were made at a constant external pH, pHo, i.e. when a reduction in [HCO3]o was accompanied by a simultaneous reduction in the partial pressure of CO2, PCO2. In contrast, if PCO2 was maintained constant, then a change in [HCO3]o (thus producing a change in pHo) resulted in an inverse change in aiCl. This change in aiCl was also accompanied by a change in pHi: when aiCl increased, pHi decreased and vice versa. The anion-exchange inhibitor, DIDS (4,4-diisothiocyanato-stilbene disulphonic acid) abolished the effect on aiCl of changes in [HCO3]o and pHo (at constant PCO2). Furthermore DIDS reduced the influence of pHo upon pHi. Both the fall of aiCl in Cl-free solution and the subsequent reuptake of Cl following re-exposure to Cl-containing solution were slowed by a reduction in [HCO3]o (constant pHo, reduced PCO2). Both reuptake and wash-out of Cl were saturating functions of [HCO3]o with half-maximal effect occurring at an [HCO3]o of 1-1.3 mmol/l. The reuptake of Cl was little affected by removal of external Na (bis,2-hydroxy ethyl, dimethyl ammonium substituted). The reuptake of Cl was unaffected by amiloride (1 mmol/l) but slowed by piretanide (1 mmol/l).(ABSTRACT TRUNCATED AT 400 WORDS)
采用液体离子交换剂氯选择性微电极和玻璃凹尖pH选择性微电极,在分离的绵羊心脏浦肯野纤维中测量细胞内氯活性(aiCl)和细胞内pH(pHi)。去除细胞外氯(用葡萄糖醛酸替代)会使aiCl从约20 mmol/L降至约4 mmol/L:残留水平可能是由于细胞内对氯敏感电极的干扰所致。纤维重新暴露于细胞外氯水平升高的环境中,在稳态下,aiCl水平会升高。稳态aiCl对细胞外氯活性aoCl的依赖性大致呈双曲线,在aoCl约为9.5 mmol/L时恢复50%。在所有测试的细胞外氯水平下,氯积累到远高于被动电化学平衡预测的水平。将氯耗尽的纤维暴露于不同水平的细胞外氯中,aiCl会随时间呈指数上升。aiCl的初始上升速率估计是aoCl的饱和函数,在aoCl约为33 mmol/L时出现半最大效应。上升速率比使用已发表的氯渗透系数PCl值根据恒定场理论预测的速率大约高10倍。如果在恒定的细胞外pH(pHo)下改变细胞外碳酸氢盐浓度[HCO3]o,稳态aiCl基本上对其变化不敏感,即当[HCO3]o降低同时二氧化碳分压PCO2也降低时。相反,如果PCO2保持恒定,那么[HCO3]o的变化(从而导致pHo变化)会导致aiCl发生相反变化。aiCl的这种变化也伴随着pHi的变化:当aiCl增加时,pHi降低,反之亦然。阴离子交换抑制剂DIDS(4,4 - 二异硫氰酸根合芪二磺酸)消除了[HCO3]o和pHo变化(在恒定PCO2下)对aiCl的影响。此外,DIDS降低了pHo对pHi的影响。[HCO3]o降低(恒定pHo,降低PCO2)会减缓无氯溶液中aiCl的下降以及重新暴露于含氯溶液后氯的后续再摄取。氯的再摄取和洗脱都是[HCO3]o的饱和函数,在[HCO3]o为1 - 1.3 mmol/L时出现半最大效应。去除细胞外钠(用双(2 - 羟乙基)二甲基铵替代)对氯的再摄取影响不大。氯的再摄取不受氨氯地平(1 mmol/L)影响,但受吡咯他尼(1 mmol/L)减缓。(摘要截短于400字)