Differential Expression and Subcellular Localization of Claudin-7, −8, −12, −13, and −15 Along the Mouse Intestine

• DOI: 10.1369/jhc.6a6944.2006
• Publication Date: 2006-05-02T00:19:38Z
• Authors: Hiroki Fujita; Hideki Chiba; Hiroshi Yokozaki; Naoyuki Sakai; Kotaro Sugimoto; Takuro Wada; Takashi Kojima; Toshihiko Yamashita; Norimasa Sawada
• Keywords: Male; 0301 basic medicine; 0303 health sciences; Reverse Transcriptase Polymerase Chain Reaction; Immunoblotting; Fluorescent Antibody Technique; Membrane Proteins; Tight Junctions; 3. Good health; Intestines; Mice, Inbred C57BL; Mice; 03 medical and health sciences; Antibody Specificity; COS Cells; Chlorocebus aethiops; Claudins; Animals; Female; RNA, Messenger; Rabbits; Intestinal Mucosa; Subcellular Fractions

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.