Civan M M, Garty H
Methods Enzymol. 1990;192:683-97. doi: 10.1016/0076-6879(90)92102-j.
Sodium ion transport across tight epithelia has been investigated particularly extensively by studying two model systems: the urinary bladder of the toad and the frog skin. The greatest advantage presented by these models is the capability of monitoring net transepithelial Na+ flux simply, precisely, and instantaneously by measurement of the short circuit current (ISC). Many of the caveats involved in the measurement are discussed in detail. In order to fully characterize the forces driving Na+ movement across the series apical and basolateral membranes, it is necessary to measure intracellular potential and ionic composition. Such measurements are far more easily conducted with frog skin than with toad bladder, using the major biophysical techniques currently available. Regulation of transepithelial Na+ movement across tight epithelia is largely conducted at the apical membranes. This regulation can be clarified by study of the isolated Na+ channels in membrane vesicles. Such vesicles are far more easily prepared from toad urinary bladder than from frog skin. The strengths and potential misappropriations of this technique are considered in detail.
通过研究两个模型系统,即蟾蜍的膀胱和蛙皮,对钠离子跨紧密上皮的转运进行了特别广泛的研究。这些模型的最大优势在于能够通过测量短路电流(ISC)简单、精确且即时地监测跨上皮的净钠离子通量。详细讨论了测量中涉及的许多注意事项。为了全面表征驱动钠离子穿过顶端和基底外侧膜序列的力,有必要测量细胞内电位和离子组成。使用目前可用的主要生物物理技术,在蛙皮上进行此类测量比在蟾蜍膀胱上要容易得多。跨紧密上皮的上皮钠离子转运调节主要在顶端膜进行。通过研究膜囊泡中分离的钠离子通道可以阐明这种调节。从蟾蜍膀胱制备此类囊泡比从蛙皮要容易得多。详细考虑了该技术的优势和可能的不当应用。