Duncan G, Stewart S, Prescott A R, Warn R M
School of Biological Sciences, University of East Anglia, Norwich, United Kingdom.
J Membr Biol. 1988 Jun;102(3):195-204. doi: 10.1007/BF01925713.
The isolated frog lens epithelium can be maintained intact in both appearance and electrical properties for more than 24 hours. The mean resting membrane potential was -80 mV and the cells were depolarized by both high potassium and low calcium Ringer's solution in a manner very similar to that of the whole lens. The epithelial cells were found to be well coupled using both electrical and dye-injection techniques. Electrical coupling was measured using separate current-injection and voltage-measuring electrodes and the relationship between the induced voltage and distance from the current-passing electrode could be well fitted by a Bessel Function solution to the cable equation. The values obtained from the fit for the membrane and internal resistances were 1.95 omega m2 and 25 omega m, respectively. Exposure to octanol (500 microM) or low external Ca2+ (less than 1 microM) failed to disrupt significantly the intercellular flow of current. There was evidence to suggest that raised intracellular calcium does, however, uncouple the cells. Dye coupling was investigated by microinjecting Lucifer Yellow CH into single epithelial cells. Diffusion into surrounding cells was rapid and, in control medium, occurred in a radially symmetrical manner. In contrast to the electrical coupling data, dye transfer appeared to be blocked by exposure to 500 microM octanol and was severely restricted on perfusing with low external calcium. Differences between the electrical and dye-coupling experiments indicate either that there are two types of junction within the cell and only the larger type, permeable to Lucifer Yellow, is capable of being uncoupled or that there is only one large type of junction which can be partially closed by uncoupling agents.
分离出的青蛙晶状体上皮细胞在外观和电学特性上都能保持完整超过24小时。平均静息膜电位为-80mV,高钾和低钙任氏液都会使细胞去极化,其方式与整个晶状体非常相似。利用电学和染料注射技术发现上皮细胞之间耦合良好。使用单独的电流注入电极和电压测量电极来测量电耦合,诱导电位与离电流通过电极的距离之间的关系可以用电缆方程的贝塞尔函数解很好地拟合。拟合得到的膜电阻和内阻的值分别为1.95Ω·m²和25Ω。暴露于辛醇(500μM)或低细胞外Ca²⁺(小于1μM)并不会显著破坏细胞间的电流流动。然而,有证据表明细胞内钙升高确实会使细胞解耦联。通过将荧光黄CH显微注射到单个上皮细胞中来研究染料耦合。染料迅速扩散到周围细胞中,在对照培养基中,以径向对称的方式发生。与电耦合数据相反,暴露于500μM辛醇似乎会阻止染料转移,在用低细胞外钙灌注时染料转移受到严重限制。电耦合和染料耦合实验之间的差异表明,要么细胞内有两种类型的连接,只有较大的、对荧光黄可渗透的那种连接能够被解耦联,要么只有一种大的连接类型,它可以被解耦联剂部分关闭。