Reinach P S, Tarvin J T, Hirsch M
Department of Ophthalmology, Washington University School of Medicine, St. Louis, MO.
Biochim Biophys Acta. 1991 Jul 22;1066(2):115-23. doi: 10.1016/0005-2736(91)90177-a.
In the isolated bullfrog cornea, measurements of DC electrical parameters in conjunction with AC impedance and ultrastructural analyses were used to determine the effects of 10(-5) M amphotericin B on epithelial cellular membrane and paracellular conductances. In NaCl Ringers, amphotericin B elicited a 3.5-fold increase in the specific apical membrane conductance (Ga/Ca); where Ga and Ca are the apical membrane conductance and capacitance, respectively. The basolateral membrane conductance (Gb) and the basolateral membrane capacitance (Cb) fell by 57% and 50%, respectively. In the paracellular pathway, the tight junctional complex (Gj) was unchanged whereas the lateral intercellular space resistance (Rp) decreased by 55%. The declines in Gb and Cb were suggestive of cell volume shrinkage because these changes were consistent with a previously described decline in intracellular K+ content and reduction in exposed basolateral membrane area to current flow. Ultrastructural analysis validated that amphotericin B caused cell volume shrinkage because there was: (1) increased folding of the basolateral membrane and waviness of the basal aspects- of the plasma membrane; (2) dilatation of the lateral intercellular spaces. This agreement suggests that intracellular activity decreased following exposure to amphotericin B which resulted in cell volume shrinkage and an impairment of Cl- uptake across the basolateral membrane.
在分离的牛蛙角膜中,结合交流阻抗测量直流电学参数并进行超微结构分析,以确定10⁻⁵ M两性霉素B对上皮细胞膜和细胞旁电导的影响。在氯化钠林格氏液中,两性霉素B使特异性顶膜电导(Ga/Ca)增加了3.5倍;其中Ga和Ca分别是顶膜电导和电容。基底外侧膜电导(Gb)和基底外侧膜电容(Cb)分别下降了57%和50%。在细胞旁途径中,紧密连接复合体(Gj)未发生变化,而细胞间侧向间隙电阻(Rp)下降了55%。Gb和Cb的下降提示细胞体积缩小,因为这些变化与先前描述的细胞内K⁺含量下降以及暴露于电流的基底外侧膜面积减少相一致。超微结构分析证实两性霉素B导致细胞体积缩小,原因如下:(1)基底外侧膜折叠增加以及质膜基底部分出现波纹;(2)细胞间侧向间隙扩张。这种一致性表明,暴露于两性霉素B后细胞内活性降低,导致细胞体积缩小以及跨基底外侧膜的Cl⁻摄取受损。