Differential expression and subcellular localization of claudin-7, -8, -12, -13, and -15 along the mouse intestine.
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.
Fujita
Hiroki
H
Dept. of Pathology, Sapporo Medical University School of Medicine, S1, W17, Sapporo 060-8556, Japan.
Chiba
Hideki
H
Yokozaki
Hiroshi
H
Sakai
Naoyuki
N
Sugimoto
Kotaro
K
Wada
Takuro
T
Kojima
Takashi
T
Yamashita
Toshihiko
T
Sawada
Norimasa
N
D000818:Animals; D000918:Antibody Specificity; D019556:COS Cells; D002522:Chlorocebus aethiops; D057167:Claudins; D005260:Female; D005455:Fluorescent Antibody Technique; D015151:Immunoblotting; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000033:anatomy & histology; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D012333:RNA, Messenger / Q000096:biosynthesis; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013347:Subcellular Fractions / Q000378:metabolism*; D019108:Tight Junctions / Q000378:metabolism*
D057167:Claudins; C507360:Cldn7 protein, mouse; D008565:Membrane Proteins; D012333:RNA, Messenger; C488754:claudin 12 protein, mouse; C459596:claudin 15; C502011:claudin 8