Willumsen N J, Boucher R C
Department of Medicine, School of Medicine, University of North Carolina, Chapel Hill 27514.
Am J Physiol. 1989 May;256(5 Pt 1):C1054-63. doi: 10.1152/ajpcell.1989.256.5.C1054.
A method for determination of shunt resistance (Rs) and absolute conductive ion permeabilities of the apical membrane in epithelia from steady-state data is described. The method assumes that the currents are satisfactorily described by the Goldman-Hodgkin-Katz regime. Its application requires measurements of standard transepithelial electrophysiological parameters and of one or more intracellular ion activities. It is applicable under both open- and short-circuit conditions. The method was tested in an electrophysiological analysis of cultured normal and cystic fibrosis (CF) human nasal epithelium. In 15 normal and 10 CF preparations with mean transepithelial resistances of 338 and 427 omega.cm2, Rs was 412 and 623 omega.cm2, respectively. The Rs values determined with the present method were strongly correlated (r = 0.94) with those obtained with another method available in the electrophysiological literature but were as a mean 20% lower. Amiloride increased Rs by 25% in CF and by 8% in normal preparations. In normal preparations, the apical Cl permeability (PCla) was 3.6 x 10(-6) cm/s, and the apical Na permeability (PNaa) was 1.6 x 10(-6) cm/s. In CF preparations, PCla was reduced to a maximum of 2.3 x 10(-7) cm/s, whereas PNaa was increased to 6.2 x 10(-6) cm/s. The apical membrane electromotive force was -1 mV in normal and 43 mV in CF preparations. It is concluded that the method can be used to calculate Rs, apical membrane ion permeabilities, and electromotive forces from steady-state electrophysiological data.
本文描述了一种根据稳态数据测定上皮细胞顶膜分流电阻(Rs)和绝对导电离子渗透率的方法。该方法假定电流可用戈德曼-霍奇金-卡茨机制得到满意描述。其应用需要测量标准跨上皮电生理参数以及一种或多种细胞内离子活性。该方法在开路和短路条件下均适用。在对培养的正常和囊性纤维化(CF)人鼻上皮进行的电生理分析中对该方法进行了测试。在15个正常样本和10个CF样本中,平均跨上皮电阻分别为338和427Ω·cm²,Rs分别为412和623Ω·cm²。用本方法测定的Rs值与电生理文献中另一种可用方法得到的值高度相关(r = 0.94),但平均低20%。氨氯地平使CF样本中的Rs增加25%,使正常样本中的Rs增加8%。在正常样本中,顶膜Cl渗透率(PCla)为3.6×10⁻⁶cm/s,顶膜Na渗透率(PNaa)为1.6×10⁻⁶cm/s。在CF样本中,PCla降至最大2.3×10⁻⁷cm/s,而PNaa增加到6.2×10⁻⁶cm/s。正常样本中的顶膜电动势为-1mV,CF样本中的为43mV。结论是该方法可用于根据稳态电生理数据计算Rs、顶膜离子渗透率和电动势。