Ishikawa T, Cook D I
Department of Physiology, University of Sydney, NSW, Australia.
Pflugers Arch. 1994 Jun;427(3-4):203-9. doi: 10.1007/BF00374525.
We have used the single-channel patch-clamp technique to study ion channels in the plasma membrane of the HSG human submandibular gland duct cell line. In cell-attached and excised inside-out patches, at least six channel types were observed. When the pipette contained an isotonic KCl-rich solution and the bath an isotonic NaCl-rich solution, the predominant channel type seen in excised inside-out patches was a Cl- channel with an outwardly rectifying current/voltage (I/V) relation that had a conductance of 12 pS at positive pipette potentials and 43 pS at negative pipette potentials. The channel was only seen in excised patches and its open probability was not significantly increased by membrane depolarization. The channel selectivity sequence (relative to Cl-) was estimated from reversal potential measurements to be: SCN- (1.8) > NO3- (1.4) > I- (1.1) approximately Cl- (1) approximately Br- (0.8) > acetate (0.35). In inside-out patches the channel was blocked by addition of 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB) (100 mumol/l) to the bath but not by 9-anthracene carboxylic acid (9-AC) (100 mumol/l). The channel was not activated by increases in the free Ca2+ concentration on the cytosolic surface. This is the first report of an outwardly rectifying Cl- channel in a salivary epithelium. The properties of this channel are not in accordance with the properties of the Cl- conductances in the acinar or duct tissues which have been studied so far and its physiological role is unclear.
我们运用单通道膜片钳技术研究了HSG人下颌下腺导管细胞系质膜中的离子通道。在细胞贴附式和内面向外式膜片中,至少观察到了六种通道类型。当移液管中含有富含KCl的等渗溶液,而浴槽中含有富含NaCl的等渗溶液时,在内面向外式膜片中观察到的主要通道类型是一种Cl-通道,其电流/电压(I/V)关系呈外向整流,在移液管正电位时电导为12 pS,在移液管负电位时电导为43 pS。该通道仅在内面向外式膜片中可见,其开放概率不会因膜去极化而显著增加。通过反转电位测量估计,通道选择性序列(相对于Cl-)为:SCN-(1.8)> NO3-(1.4)> I-(1.1)≈ Cl-(1)≈ Br-(0.8)> 乙酸盐(0.35)。在内面向外式膜片中,向浴槽中添加5-硝基-2-(3-苯丙基氨基)-苯甲酸(NPPB)(100 μmol/L)可阻断该通道,但添加9-蒽甲酸(9-AC)(100 μmol/L)则不能。该通道不会因胞质表面游离Ca2+浓度的增加而被激活。这是唾液上皮细胞中外向整流Cl-通道的首次报道。该通道的特性与迄今为止所研究的腺泡或导管组织中的Cl-电导特性不符,其生理作用尚不清楚。