Alimentary TractHeterogeneity in expression and subcellular localization of claudins 2, 3, 4, and 5 in the rat liver, pancreas, and gut☆,☆☆
Section snippets
Tissue preparation
Male Sprague–Dawley rats (200–250 g) were obtained through approved protocols of the Yale Liver Center from the Yale Animal Resources Center. Anesthesia was administered by intraperitoneal injection of pentobarbital (5 mg/100 g body wt; Abbott Laboratories, North Chicago, IL). Rats were anesthetized with pentobarbital and underwent laparotomy, and the distal abdominal aorta was exposed and cannulated with a polyethylene-90 catheter. Organs were systemically rinsed free of blood with ice-cold
Specificity of anticlaudin antibodies
To investigate the distribution and localization of claudins in the gastrointestinal tract, we generated rabbit polyclonal antibodies against peptides corresponding to the C-terminal sequences of claudins 1 through 5. Either rat, mouse, or human sequences were used depending on which were available in the database (see Materials and Methods). Subsequent availability of all mouse and human sequences shows them to be conserved and suggests that antibodies raised against these sequences will also
Discussion
In this study, we have documented the tissue distribution and subcellular localization of claudins 2–5 in the gastrointestinal tract. Earlier Northern analysis had indicated the heterogeneity and possible tissue and cell type specificity in expression of some claudins.3, 13, 14, 15, 16 Two claudins had already been shown to have extremely restricted expression patterns: claudin 16 in the thick ascending loop of Henle16 and claudin 11 in Sertoli cells and oligodendrocytes.14, 15 We observed
Acknowledgements
The authors thank Drs. Christina M. Van Itallie, Alan Fanning, and Zenta Walther for helpful discussions and Keith Choate and Dr. Richard Lifton for providing a paracellin-GST fusion protein.
References (32)
- et al.
CNS myelin and sertoli cell tight junction strands are absent in Osp/claudin-11 null mice
Cell
(1999) - et al.
Claudin-1 contributes to the epithelial barrier function in MDCK cells
Eur J Cell Biol
(1999) - et al.
The tight junction protein ZO-1 establishes a link between the transmembrane protein occludin and the actin cytoskeleton
J Biol Chem
(1998) - et al.
Tight junction proteins ZO-1, ZO-2, and occludin along isolated renal tubules
Kidney Int
(2000) - et al.
Structural changes in the plasma membrane accompanying differentiation of epithelial cells in human and monkey small intestine
Gastroenterology
(1980) - et al.
Localization and function of the organic anion-transporting polypeptide Oatp2 in rat liver
Gastroenterology
(1999) - et al.
Claudins regulate the intestinal barrier in response to immune mediators
Gastroenterology
(2000) Heterogeneity of intestinal transport
Digestion
(1998)Barrier function of epithelia
Am J Physiol
(1981)- et al.
Claudin-1 and -2: novel integral membrane proteins localizing at tight junctions with no sequence similarity to occludin
J Cell Biol
(1998)
Claudin multigene family encoding four-transmembrane domain protein components of tight junction strands
Proc Natl Acad Sci U S A
Morphological factors influencing transepithelial per-meability: a model for the resistance of the zonula occludens
J Membr Biol
Molecular architecture of tight junctions
Annu Rev Physiol
Freeze-fracture replica electron microscopy combined with SDS digestion for cytochemical labeling of integral membrane proteins. Application to the immunogold labeling of intra-cellular junctional complexes
J Cell Sci
Occludin confers adhesiveness when expressed in fibroblasts
J Cell Sci
Ca(2+)-independent cell-adhesion activity of claudins, a family of integral membrane proteins localized at tight junctions
Curr Biol
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Address requests for reprints to: Christoph Rahner, M.D., Section of Digestive Diseases, 1080 LMP, Department of Internal Medicine, Yale School of Medicine, 333 Cedar Street, New Haven, Connecticut 06520-8019. e-mail: [email protected]; fax: (203) 785-7273.
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Supported by National Institutes of Health grants (DK45134 to J.M.A.) and the Core Facilities of the Yale Liver Center (DK34989).