Ishikawa T, Cook D I
Department of Physiology, University of Sydney, N.S.W., Australia.
J Membr Biol. 1993 Sep;135(3):261-71. doi: 10.1007/BF00211098.
Previous studies have shown that the whole-cell current-voltage (I-V) relation of unstimulated sheep parotid cells is dominated by two K+ conductances, one outwardly and the other inwardly rectifying. We now show that once these K+ conductances are blocked by replacement of pipette K+ with Na+ and by the addition of 5 mmol/liter CsCl to the bath, there remains an outwardly rectifying conductance with a reversal potential of 0 mV. Replacement of 120 mmol/liter NaCl in the pipette solution with an equimolar amount of Na-glutamate shifted the reversal potential of this residual current to -55 mV, indicating that the conductance was Cl- selective. The Cl- current was activated by increasing the free Ca2+ in the pipette solution from 10 to 100 nmol/liter. When the Ca2+ concentration in the pipette solution was 10 nmol/liter, the relaxations observed in response to membrane depolarization could be fitted with a single exponential, whose time constant increased from 81 to 183 ms as the pipette potential was increased from -30 to +60 mV. Relaxation analysis showed that the current was activated by membrane depolarization. Reversal potential measurements in experiments in which external Cl- was replaced with various anions, gave the following relative permeabilities: SCN- (1.80) > I- (1.09) > Cl- (1) > NO3- (0.92) > Br- (0.75). The relative conductances were: SCN- (2.18) > I- (1.07) > Cl- (1.00) > Br- (0.91) > NO3- (0.50). The Cl- current was blocked by NPPB (ID50 approximately 10 microM), DIDS (10 or 30 mumol/liter) and furosemide (100 mumol/liter).
先前的研究表明,未受刺激的绵羊腮腺细胞的全细胞电流-电压(I-V)关系主要由两种钾离子电导主导,一种是外向整流,另一种是内向整流。我们现在表明,一旦这些钾离子电导被用钠离子替代移液管中的钾离子以及向浴液中添加5 mmol/升氯化铯所阻断,仍然存在一种外向整流电导,其反转电位为0 mV。用等摩尔量的谷氨酸钠替代移液管溶液中的120 mmol/升氯化钠,使这种残余电流的反转电位变为-55 mV,表明该电导对氯离子具有选择性。通过将移液管溶液中的游离钙离子从10 nmol/升增加到100 nmol/升,氯离子电流被激活。当移液管溶液中的钙离子浓度为10 nmol/升时,响应膜去极化所观察到的弛豫可以用单一指数拟合,随着移液管电位从-30 mV增加到+60 mV,其时间常数从81 ms增加到183 ms。弛豫分析表明,电流是由膜去极化激活的。在将外部氯离子替换为各种阴离子的实验中进行的反转电位测量,得出以下相对渗透率:硫氰酸根离子(1.80)>碘离子(1.09)>氯离子(1)>硝酸根离子(0.92)>溴离子(0.75)。相对电导率为:硫氰酸根离子(2.18)>碘离子(1.07)>氯离子(1.00)>溴离子(0.91)>硝酸根离子(0.50)。氯离子电流被NPPB(半数抑制浓度约为10 μM)、DIDS(10或30 μmol/升)和呋塞米(100 μmol/升)阻断。