Kroll B, Bremer S, Tümmler B, Kottra G, Frömter E
Zentrum der Physiologie, Klinikum der J.W. Goethe-Universität, Frankfurt/Main, Federal Republic of Germany.
Pflugers Arch. 1991 Aug;419(1):101-7. doi: 10.1007/BF00373753.
The epithelial Na+ conductance was expressed in Xenopus laevis oocytes by injection of size-fractionated mRNA of bovine tracheal epithelium. Fractionation was achieved by sucrose density gradient centrifugation. Successful expression was analysed by recording current/voltage (I/V) curves in the presence and absence of amiloride (10 mumol/l). The newly expressed conductance was half-maximally inhibited by 44 nmol/l amiloride and exhibited a selectivity for Na+ over K+ of 140:1. I/V curves obtained at different extracellular Na+ concentrations ([Na+]o) were subjected to a Goldman-fit analysis to obtain the relation between Na+ permeability (PNa) and [Na+]o. The data show that decreasing [Na+]o from 85 mmol/l to 0.85 mmol/l increased PNa by more than threefold, which is thought to reflect Na+ channel inhibition by increasing [Na+]o. This effect clearly exceeded what can be attributed to concentration saturation of single Na+ channel conductance (Palmer and Frindt (1986) Proc Natl Acad Sci USA 83:2767). No correlation of inhibition with intracellular Na+ concentration was observed. Preservation of the [Na+]o-dependent self-inhibition by the newly expressed Na+ conductance suggests that it is an intrinsic property of the Na+ channel protein, probably mediated by an extracellular Na+ binding site.
通过注射经大小分级的牛气管上皮mRNA,在非洲爪蟾卵母细胞中表达上皮钠电导。分级分离通过蔗糖密度梯度离心实现。在存在和不存在氨氯吡咪(10 μmol/L)的情况下记录电流/电压(I/V)曲线,以分析成功表达情况。新表达的电导被44 nmol/L氨氯吡咪半最大抑制,并且对Na⁺的选择性高于K⁺,为140:1。对在不同细胞外Na⁺浓度([Na⁺]o)下获得的I/V曲线进行戈德曼拟合分析,以获得Na⁺通透性(PNa)与[Na⁺]o之间的关系。数据表明,将[Na⁺]o从85 mmol/L降至0.85 mmol/L可使PNa增加三倍以上,这被认为反映了通过增加[Na⁺]o对Na⁺通道的抑制作用。这种效应明显超过了可归因于单个Na⁺通道电导浓度饱和的程度(Palmer和Frindt(1986年),《美国国家科学院院刊》83:2767)。未观察到抑制作用与细胞内Na⁺浓度之间的相关性。新表达的Na⁺电导对[Na⁺]o依赖性自我抑制的保留表明,这是Na⁺通道蛋白的一种内在特性,可能由细胞外Na⁺结合位点介导。