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首页> 外文期刊>American Journal of Physiology >Calcium-switch technique and junctional permeability in native rabbit esophageal epithelium.
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Calcium-switch technique and junctional permeability in native rabbit esophageal epithelium.

机译：钙转换技术和天然兔食管上皮细胞的结合通透性。

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The Ca(2+)-switch technique was used to investigate the nature of the barrier governing (paracellular) permeability across the junctions of "native" rabbit esophageal epithelium. This was done by mounting esophageal epithelium in Ussing chambers to monitor transepithelial electrical resistance (R(T)), a marker of junctional permeability. When exposed to Ca(2+)-free Ringer solutions containing EDTA, R(T) declined approximately 35% below baseline over 2 h, and this decline reversed within 2 h by restoration of (1.2 mM) Ca(2+)-containing, normal Ringer solution ("Ca(2+)-switch technique"). Junctional resealing, i.e., increased R(T) on Ca(2+) replacement, was assessed by the Ca(2+)-switch technique and shown to be 1) specific for Ca(2+), with only Mn(2+) among substituted divalent cations yielding partial resealing; 2) a function of extracellular Ca(2+) levels because maneuvers (BAPTA/AM or A23187 exposure) to alter intracellular Ca(2+) had no effect; 3) dose dependent, requiring as a minimum > or =0.5 mM Ca(2+) and 1.2 mM Ca(2+) for optimization; and 4) independent of protein synthesis because it was not inhibited by cycloheximide. Resealing was also inhibited by luminal antibodies or synthetic peptides to the extracellular domain of E-cadherin. Immunohistochemistry revealed E-cadherin within all layers of stratum corneum in Ca(2+)-free but not Ca(2+)-containing solution. The present investigation documents, using the Ca(2+)-switch technique, that esophageal epithelial junctions contain a major Ca(2+)-dependent component and that this component reflects adhesion between the extracellular domains of E-cadherin containing a histidine-alanine-valine recognition sequence.
机译：Ca（2+）开关技术用于研究控制“天然”兔食管上皮的交界处的屏障（细胞旁）通透性的屏障的性质。这是通过将食管上皮安装在Usssing室中以监测跨上皮电阻（R（T））来实现的，该电阻是连接通透性的标志。当暴露于含有EDTA的不含Ca（2+）的林格溶液时，R（T）在2小时内下降至比基线低约35％，并且该下降在2小时内通过恢复（1.2 mM）含Ca（2+）的作用而逆转，正常的林格溶液（“ Ca（2 +）-switch technique”）。交界处重新密封，即增加Ca（2+）的R（T），通过Ca（2+）转换技术进行评估，并显示为1）Ca（2+）特有，只有Mn（2+） ）在取代的二价阳离子中产生部分重密封; 2）细胞外Ca（2+）水平的函数，因为通过操纵（BAPTA / AM或A23187暴露）来改变细胞内Ca（2+）没有作用; 3）剂量依赖性，需要至少> 0.5mM Ca（2+）和1.2 mM Ca（2+）进行优化; 4）独立于蛋白质合成，因为它不受环己酰亚胺的抑制。再密封也受到针对E-钙粘着蛋白细胞外结构域的腔抗体或合成肽的抑制。免疫组织化学揭示了在不含Ca（2+）但不含Ca（2+）的溶液中角质层所有层中的E-cadherin。本研究文件使用Ca（2+）转换技术，证明食管上皮连接处含有主要的Ca（2+）依赖性成分，并且该成分反映了含有组氨酸-丙氨酸的E-钙粘着蛋白胞外域之间的粘附-缬氨酸识别序列。

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