Bukauskas F F, Kempf C, Weingart R
Department of Physiology, Bern, Switzerland.
Exp Physiol. 1992 Nov;77(6):903-11. doi: 10.1113/expphysiol.1992.sp003657.
Cell pairs of an insect cell line (Aedes albopictus, clone C6/36) were used study simultaneously the diffusional and electrical properties of intercellular junctions. Diffusion studies involved injection of fluorescent molecules into one cell of a cell pair and visual inspection of their intercellular redistribution. Electrical measurements involved a dual voltage clamp method and whole-cell recording with patch pipette. The voltage clamp protocol was aimed at examining the dependency of the junctional conductance, gj, on membrane potential, Vm. Cell pairs exhibiting a voltage-dependent gj were found to allow intercellular diffusion of Lucifer Yellow CH (molecular mass, 443 Da), but not of FITC-dextran (molecular mass, 4,400 Da). This response pattern is consistent with the presence of gap junctions in the intercellular junctions. Cell pairs showing no voltage dependence of gj were found to permit intercellular diffusion of both Lucifer Yellow CH and FITC-dextran (dextran labelled with fluorescein isothiocyanate). This behaviour is compatible with the presence of cytoplasmic bridges connecting the two adjacent cells. Hence, in culture the cells investigated express two kinds of intercellular structures, gap junctions and cytoplasmic bridges.
利用一种昆虫细胞系(白纹伊蚊,克隆C6/36)的细胞对同时研究细胞间连接的扩散特性和电学特性。扩散研究包括将荧光分子注入细胞对中的一个细胞,并目视检查其在细胞间的重新分布。电学测量采用双电压钳法和膜片吸管全细胞记录。电压钳实验方案旨在研究连接电导gj对膜电位Vm的依赖性。发现表现出电压依赖性gj的细胞对允许荧光黄CH(分子量443道尔顿)在细胞间扩散,但不允许异硫氰酸荧光素标记的葡聚糖(分子量4400道尔顿)扩散。这种反应模式与细胞间连接中存在间隙连接一致。发现gj无电压依赖性的细胞对允许荧光黄CH和异硫氰酸荧光素标记的葡聚糖在细胞间扩散。这种行为与连接两个相邻细胞的胞质桥的存在相符。因此,在培养中,所研究的细胞表达两种细胞间结构,即间隙连接和胞质桥。