Kunzelmann K, Gerlach L, Fröbe U, Greger R
Physiologisches Institut, Albert-Ludwigs-Universität Freiburg, FederalRepublic of Germany.
Pflugers Arch. 1991 Feb;417(6):616-21. doi: 10.1007/BF00372960.
Bicarbonate permeability of epithelial chloride channels has been studied using the patch-clamp technique. The experiments were performed in excised inside-out oriented membrane patches from three different cultured cell types: (a) HT29 colon carcinoma cell line, (b) T84 colon carcinoma cell line, and (c) respiratory epithelial cells (REC) in primary culture. In all three preparations we observed outwardly rectifying chloride channels with similar conductances with 145 mmol/l NaCl solution in the pipette and in the bath (Cl- pipette/Cl- bath). When Cl- was replaced by HCO3- in the bath (Cl-/HCO3-) the conductance of the channel at negative clamp voltages was reduced significantly by 40% for HT29 (n = 6), 39% for T84 (n = 7), and 38% for REC (n = 6). Similarly, the zero-current potential (V1 = 0) was shifted from 0 mV (Cl-/Cl-) to negative values (Cl-/HCO3-) revealing permeability ratios PCl/PHCO3 of 2.4 +/- 0.1 for HT29 (n = 6), 2.0 +/- 0.1 for T84 (n = 7), and 1.8 +/- 0.1 for REC (n = 7). With NaHCO3 as the pipette solution and NaCl in the bath, the V1 = 0 was positive and a PCl/PHCO3 value of 2.3 +/- 0.1 was determined for HT29 (n = 5). Replacement of Cl- in the bath by HCO3- reduced V1 = 0 to values close to 0 mV. In another series of experiments, the pipette was filled with 145 mmol/l NaCl and the bath contained 35 mmol/l NaCl to which 35 mmol/l NaHCO3 were added. We found that neither the conductance for the inward current nor V1 = 0 was changed significantly with the addition of NaHCO3 (HT29, n = 6). We conclude that the HCO3- permeability and HCO3- conductance of these channels is about half of that for Cl-.
利用膜片钳技术研究了上皮氯离子通道的碳酸氢根通透性。实验在三种不同培养细胞类型的外翻式膜片中进行:(a)HT29结肠癌细胞系,(b)T84结肠癌细胞系,以及(c)原代培养的呼吸道上皮细胞(REC)。在所有这三种制剂中,我们观察到外向整流氯离子通道,在移液管和浴槽中使用145 mmol/l NaCl溶液时具有相似的电导(Cl-移液管/Cl-浴槽)。当浴槽中的Cl-被HCO3-取代(Cl-/HCO3-)时,对于HT29(n = 6),负钳制电压下通道的电导显著降低40%;对于T84(n = 7),降低39%;对于REC(n = 6),降低38%。同样,零电流电位(V1 = 0)从0 mV(Cl-/Cl-)移至负值(Cl-/HCO3-),显示HT29的PCl/PHCO3为2.4±0.1(n = 6),T84为2.0±0.1(n = 7),REC为1.8±0.1(n = 7)。以NaHCO3作为移液管溶液且浴槽中为NaCl时,V1 = 0为正值,HT29的PCl/PHCO3值为2.3±0.1(n = 5)。浴槽中的Cl-被HCO3-取代后,V1 = 0降至接近0 mV的值。在另一系列实验中,移液管填充145 mmol/l NaCl,浴槽含有35 mmol/l NaCl并添加35 mmol/l NaHCO3。我们发现添加NaHCO3后内向电流的电导和V1 = 0均未显著改变(HT29,n = 6)。我们得出结论,这些通道的HCO3-通透性和HCO3-电导约为Cl-的一半。