Hirsch J, Schlatter E
Albert-Ludwigs-Universität, Physiologisches Institut, Freiburg, Germany.
Pflugers Arch. 1993 Sep;424(5-6):470-7. doi: 10.1007/BF00374910.
Impalement studies in isolated perfused cortical collecting ducts (CCD) of rats have shown that the basolateral membrane possesses a K+ conductive pathway. In the present study this pathway was investigated at the single-channel level using the patch-clamp technique. Patch-clamp recordings were obtained from enzymatically isolated CCD segments and freshly isolated CCD cells with the conventional cell-free, cell-attached and the cell-attached nystatin method. Two K+ channels were found which were highly active on the cell with a conductance of 67 +/- 5 pS (n = 18) and 148 +/- 4 pS (n = 21) with 145 mmol/l K+ in the pipette. In excised patches the first channel had a conductance of 28 +/- 2 pS (n = 15), whereas the second one had a conductance of 85 +/- 1 pS (n = 53) at 0 mV clamp voltage with 145 mmol/l K+ on one side and 3.6 mmol/l K+ on the other side of the membrane. So far it has not been possible to characterize the smaller channel further. Excised, and with symmetrical K+ concentrations of 145 mmol/l, the intermediate channel had a linear conductance of 198 +/- 19 pS (n = 5). After excision in the inside-out configuration the open probability (Po) of this channel was low (0.18 +/- 0.05, n = 13) whereas in the outside-out configuration this channel had a threefold higher Po (0.57 +/- 0.04, n = 12). Several inhibitors were tested in excised membranes. Ba2+ (1 mmol/l), tetraethylammonium (TEA+, 10 mmol/l) and verapamil (0.1 mmol/l) all blocked this channel reversibly.(ABSTRACT TRUNCATED AT 250 WORDS)
对大鼠离体灌注皮质集合管(CCD)进行的刺入研究表明,基底外侧膜具有钾离子传导途径。在本研究中,使用膜片钳技术在单通道水平对该途径进行了研究。通过传统的无细胞、细胞贴附和细胞贴附制霉菌素法,从酶解分离的CCD节段和新鲜分离的CCD细胞中获得膜片钳记录。发现了两种钾离子通道,在移液管中含有145 mmol/L钾离子时,这两种通道在细胞上具有高活性,电导分别为67±5 pS(n = 18)和148±4 pS(n = 21)。在切除的膜片中,在钳制电压为0 mV、膜一侧为145 mmol/L钾离子而另一侧为3.6 mmol/L钾离子时,第一种通道的电导为