McNicholas C M, Brown C D, Turnberg L A
Epithelial Membrane Research Centre, Hope Hospital, Salford, United Kingdom.
Am J Physiol. 1994 Dec;267(6 Pt 1):G1004-11. doi: 10.1152/ajpgi.1994.267.6.G1004.
The aim of the present study was to examine the possibility that Cl- uptake into both villus and crypt epithelial cells of rat duodenum occurs via an electroneutral Na-K-Cl coupled-transport mechanism. Sheets of villus cells and whole crypts were isolated using a Ca2+ chelation technique combined with continuous vibration at low temperatures. Structurally intact, viable epithelia from defined regions along the villus-crypt axis were produced. Uptake of 86Rb+ (as a proxy for K+) into both villus and crypt cells appeared to depend on a coupled process, as evidenced by the inhibition of 86Rb+ uptake by bumetanide and by the removal of either Na+ or Cl- from the bathing media. We report an improved method of isolation of viable enterocytes from defined regions along the villus-crypt axis. We demonstrate the presence of Na-K-Cl cotransport in both villus and crypt duodenal enterocytes.
本研究的目的是检验大鼠十二指肠绒毛和隐窝上皮细胞摄取氯离子是否通过电中性的钠 - 钾 - 氯协同转运机制。采用钙离子螯合技术结合低温连续振动的方法分离绒毛细胞片层和完整的隐窝。制备出沿绒毛 - 隐窝轴特定区域结构完整、有活力的上皮细胞。86Rb +（作为钾的替代物）进入绒毛和隐窝细胞的摄取似乎依赖于一个耦联过程，布美他尼对86Rb +摄取的抑制以及从浴液中去除钠离子或氯离子均证明了这一点。我们报告了一种从沿绒毛 - 隐窝轴特定区域分离有活力肠上皮细胞的改良方法。我们证明了十二指肠绒毛和隐窝肠上皮细胞中存在钠 - 钾 - 氯共转运。
