Fujita Hiroki, Chiba Hideki, Yokozaki Hiroshi, Sakai Naoyuki, Sugimoto Kotaro, Wada Takuro, Kojima Takashi, Yamashita Toshihiko, Sawada Norimasa
Dept. of Pathology, Sapporo Medical University School of Medicine, S1, W17, Sapporo 060-8556, Japan.
J Histochem Cytochem. 2006 Aug;54(8):933-44. doi: 10.1369/jhc.6A6944.2006. Epub 2006 May 1.
Among tight-junction proteins, claudins, which play a key role in paracellular transport across epithelia, claudins 1 to 5 are expressed in the intestine, and changes in their abundance and/or distribution are considered to contribute to various gastrointestinal diseases. We investigated, by reverse transcription-PCR, immunoblot, and immunofluorescence analyses, which other claudin species were expressed in the mouse intestine, and whether they showed unique expression profiles. Rabbit polyclonal antibodies against mouse claudin-8, claudin-12, and claudin-15 were generated, and their specificity was verified by immunoblotting using COS-7 cells transfected with individual claudin cDNAs. Claudin-7, -8, -12, -13, and -15 appeared to be expressed in the duodenum, jejunum, ileum, and/or colon with remarkable variations in the expression levels along the intestinal tract, and had distinct subcellular localization in the intestinal epithelium. In addition, claudin-13 and -15 exhibited gradients along the crypt-surface axis of the colon. By contrast, claudin-6, -9, -10, -11, -14, -16, -18, and -19 were not observed in the intestine. Our results indicate that five additional species of claudins have very complex expression patterns along and within the intestine, and that this may reflect differences in paracellular permeable properties, providing valuable resources for studying the significance of these claudins in gastrointestinal disorders. This manuscript contains online supplemental material available at http://www.jhc.org. Please visit this article online to view these materials.
在紧密连接蛋白中，claudins在上皮细胞旁细胞转运中起关键作用，claudins 1至5在肠道中表达，其丰度和/或分布的变化被认为与各种胃肠道疾病有关。我们通过逆转录聚合酶链反应、免疫印迹和免疫荧光分析，研究了小鼠肠道中还表达哪些其他claudin种类，以及它们是否呈现独特的表达谱。制备了针对小鼠claudin - 8、claudin - 12和claudin - 15的兔多克隆抗体，并通过使用转染了单个claudin cDNA的COS - 7细胞进行免疫印迹验证了其特异性。Claudin - 7、- 8、- 12、- 13和- 15似乎在十二指肠、空肠、回肠和/或结肠中表达，沿肠道的表达水平有显著差异，并且在肠道上皮细胞中有不同的亚细胞定位。此外，claudin - 13和- 15沿结肠隐窝-表面轴呈现梯度分布。相比之下，在肠道中未观察到claudin - 6、- 9、- 10、- 11、- 14、- 16、- 18和- 第十九。我们的结果表明，另外五种claudin在肠道内和沿肠道具有非常复杂的表达模式，这可能反映了细胞旁通透性的差异，为研究这些claudin在胃肠道疾病中的意义提供了有价值的资源。本文包含可在http://www.jhc.org获取的在线补充材料。请在线访问本文以查看这些材料。
