Macknight A D, Civan M M, Leaf A
J Membr Biol. 1975;20(3-4):365-67. doi: 10.1007/BF01870644.
The sodium which equilibrates with 24-Na in epithelial cells of toad urinary bladders has been determined. With sodium Ringer's bathing both mucosal and serosal surfaces, 24-Na in the mucosal medium equilibrated with about 35 mmoles cellular sodium/kg cellular dry weight, representing about 20% of the total cellular sodium determined flame photometrically; 24-Na in the serosal medium equilibrated with 120 mmoles cellular sodium/kg cellular dry weight, about 80% of the total cellular sodium. With 24-Na in both media all cellular sodium was labeled within 30 min. In the absence of serosal sodium, total cellular sodium and that sodium which equilibrated with mucosal 24-Na in sodium Ringer's were both similar to the cellular sodium of mucosal origin which had been determined in epithelial cells exposed on both surfaces to sodium Ringer's. Sodium-free mucosal medium, and sodium Ringer's containing amiloride 10-4 or 10-3 M in the mucosal medium, both virtually completely inhibited transepithelial sodium transport. But, whereas the cellular sodium of mucosal origin fell to only 2 mmoles/kg cellular dry weight with sodium-free mucosal medium, an appreciable labeling of cellular sodium was found whether amiloride was present before, or only after, exposure of tissue to mucosal 24-Na. Rapid washing of the mucosal surface of hemibladders just before removal of epithelial cells for analysis removed most of this sodium labeled in the presence of amiloride, suggesting that the cellular sodium of mucosal origin consists of at least two fractions with only about two-thirds truly intracellular. The sodium transport pool measured directly in these experiments is appreciably smaller than any previous estimates of pool size all of which have been obtained by indirect techniques involving use of whole hemibladders rather than epithelial cells alone.
已测定蟾蜍膀胱上皮细胞中与24-Na平衡的钠含量。用含钠林格液同时灌注黏膜面和浆膜面时,黏膜介质中的24-Na与约35 mmol细胞钠/千克细胞干重达到平衡,占火焰光度法测定的细胞总钠量的约20%;浆膜介质中的24-Na与120 mmol细胞钠/千克细胞干重达到平衡,约占细胞总钠量的80%。当两种介质中都有24-Na时,所有细胞钠在30分钟内都被标记。在没有浆膜钠的情况下,细胞总钠量以及在含钠林格液中与黏膜24-Na平衡的钠量,都与在两面都暴露于含钠林格液的上皮细胞中测定的源自黏膜的细胞钠量相似。无钠的黏膜介质以及黏膜介质中含10-4或10-3 M氨氯吡咪的含钠林格液,几乎完全抑制了跨上皮钠转运。但是,尽管无钠的黏膜介质使源自黏膜的细胞钠量降至仅2 mmol/千克细胞干重,但无论氨氯吡咪是在组织暴露于黏膜24-Na之前还是之后存在,都发现细胞钠有明显的标记。在取出上皮细胞进行分析之前,快速冲洗半膀胱的黏膜表面,去除了大部分在氨氯吡咪存在时标记的钠,这表明源自黏膜的细胞钠至少由两部分组成,其中只有约三分之二是真正的细胞内钠。在这些实验中直接测量的钠转运池明显小于以前通过涉及使用整个半膀胱而非仅上皮细胞的间接技术获得的任何池大小估计值。