Vanheel B, de Hemptinne A, Leusen I
Laboratory of Normal and Pathological Physiology, University of Gent, Belgium.
J Mol Cell Cardiol. 1988 Mar;20(3):225-38. doi: 10.1016/s0022-2828(88)80055-5.
Isolated sheep cardiac Purkinje fibers were pulled through a latex membrane and both segments were independently superfused with modified Tyrode solutions. Transmembrane potentials and intracellular pH (pHi) were continuously measured in one segment (test compartment, TC), using double-barreled pH sensitive glass microelectrodes, while the internal H+ activity was altered in the adjacent part of the fiber (experimental compartment, EC). In the latter local pHi changes were produced by removal of CO2/HCO3-, by superfusing acidic (pH 6.8) solution, and by addition and subsequent withdrawal of NH4Cl. Withdrawal of CO2/HCO3- in EC was found to have no influence on pHi in TC at 0.35 to 1.0 mm distance. The action potential first shortened and later on prolonged above control duration after switching to HCO3- -free medium. Perfusing EC with an acidic solution had virtually no effect on pHi in TC while action potential duration (APD) increased. Addition and withdrawal of NH4+ in EC decreased, respectively increased, APD. In TC no change in pHi was observed at 0.8 to 1.0 mm distance. At shorter distance a slow acidification was seen, associated with the presence of NH4+ in EC. The presence of amiloride, a blocker of the pHi regulating mechanism, could not unmask a larger pHi change. It is concluded that, in cardiac muscle, large gradients of pHi are possible over a relatively short distance, while electrotonic interaction can produce changes in the time course of the action potential in neighbouring cells having a normal intracellular pH.
将分离的绵羊心脏浦肯野纤维穿过乳胶膜,两段纤维分别用改良的台氏液进行灌流。使用双管pH敏感玻璃微电极在一段纤维(测试室,TC)中连续测量跨膜电位和细胞内pH(pHi),同时在纤维的相邻部分(实验室,EC)改变内部H⁺活性。在后者中,通过去除CO₂/HCO₃⁻、灌流酸性(pH 6.8)溶液以及添加和随后去除NH₄Cl来产生局部pHi变化。发现在0.35至1.0毫米距离处,EC中CO₂/HCO₃⁻的去除对TC中的pHi没有影响。切换到无HCO₃⁻培养基后,动作电位首先缩短,随后延长至超过对照持续时间。用酸性溶液灌流EC对TC中的pHi几乎没有影响,而动作电位持续时间(APD)增加。EC中添加和去除NH₄⁺分别降低和增加了APD。在0.8至1.0毫米距离处,TC中的pHi没有变化。在较短距离处,观察到缓慢酸化,这与EC中存在NH₄⁺有关。氨氯吡咪(一种pHi调节机制的阻滞剂)的存在并不能揭示更大的pHi变化。得出的结论是,在心肌中,在相对较短的距离内可能存在较大的pHi梯度,而电紧张相互作用可在细胞内pH正常的相邻细胞中产生动作电位时程的变化。