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Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells.

Morita K, Sasaki H, Furuse M, Tsukita S - J. Cell Biol. (1999)

Bottom Line: In contrast, in the kidney, the claudin-5/TMVCF signal was restricted to endothelial cells of arteries, but was undetectable in those of veins and capillaries.Furthermore, when claudin-5/TMVCF cDNA was introduced into mouse L fibroblasts, TJ strands were reconstituted that resembled those in endothelial cells in vivo, i.e., the extracellular face-associated TJs.These findings indicated that claudin-5/TMVCF is an endothelial cell-specific component of TJ strands.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan.

ABSTRACT
Tight junctions (TJs) in endothelial cells are thought to determine vascular permeability. Recently, claudin-1 to -15 were identified as major components of TJ strands. Among these, claudin-5 (also called transmembrane protein deleted in velo-cardio-facial syndrome [TMVCF]) was expressed ubiquitously, even in organs lacking epithelial tissues, suggesting the possible involvement of this claudin species in endothelial TJs. We then obtained a claudin-6-specific polyclonal antibody and a polyclonal antibody that recognized both claudin-5/TMVCF and claudin-6. In the brain and lung, immunofluorescence microscopy with these polyclonal antibodies showed that claudin-5/TMVCF was exclusively concentrated at cell-cell borders of endothelial cells of all segments of blood vessels, but not at those of epithelial cells. Immunoreplica electron microscopy revealed that claudin-5/TMVCF was a component of TJ strands. In contrast, in the kidney, the claudin-5/TMVCF signal was restricted to endothelial cells of arteries, but was undetectable in those of veins and capillaries. In addition, in all other tissues we examined, claudin-5/TMVCF was specifically detected in endothelial cells of some segments of blood vessels, but not in epithelial cells. Furthermore, when claudin-5/TMVCF cDNA was introduced into mouse L fibroblasts, TJ strands were reconstituted that resembled those in endothelial cells in vivo, i.e., the extracellular face-associated TJs. These findings indicated that claudin-5/TMVCF is an endothelial cell-specific component of TJ strands.

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Localization of claudin-5/TMVCF in the kidney and the intestine. Frozen sections of the kidney (a–h) and the intestine (i and j) were double labeled with anti–claudin-5/6 pAb (a) and antioccludin mAb (b), anti–claudin-5/6 pAb (c) and anti–VE-cadherin mAb (d), anti–claudin-6 pAb (e) and anti–VE-cadherin mAb (f), or anti–claudin-5/6 pAb (i) and antioccludin mAb (j). Since anti–claudin-6 pAb yielded no signals in the kidney (e) and the intestine (data not shown), the anti–claudin-5/6 pAb staining can be considered to represent the distribution of claudin-5/TMVCF. Claudin-5/TMVCF appeared to be expressed in a subset of blood vessels (arrows in a and i) that were occludin-negative. Occludin was concentrated at TJs in distal tubules in the kidney (arrowheads in b) and intestinal epithelial cells (arrowheads in j). In the kidney, VE-cadherin–positive intertubular capillaries (arrowheads in d and f) and glomerular capillaries (asterisk in d and f) did not express claudin-5/TMVCF, but afferent and efferent arterioles (arrows in c and d) expressed both claudin-5/TMVCF and VE-cadherin. When transverse frozen sections of the thicker artery and vein (arrows and arrowheads, respectively, in g and h) of the kidney were stained with anti–claudin-5/6 pAb, only the artery was intensely stained (g). (h) Phase–contrast image. Bars: 40 μm for a–f (f); 70 μm in g and h (h); 40 μm in i and j (j). Cln, claudin; Cad, cadherin.
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Figure 5: Localization of claudin-5/TMVCF in the kidney and the intestine. Frozen sections of the kidney (a–h) and the intestine (i and j) were double labeled with anti–claudin-5/6 pAb (a) and antioccludin mAb (b), anti–claudin-5/6 pAb (c) and anti–VE-cadherin mAb (d), anti–claudin-6 pAb (e) and anti–VE-cadherin mAb (f), or anti–claudin-5/6 pAb (i) and antioccludin mAb (j). Since anti–claudin-6 pAb yielded no signals in the kidney (e) and the intestine (data not shown), the anti–claudin-5/6 pAb staining can be considered to represent the distribution of claudin-5/TMVCF. Claudin-5/TMVCF appeared to be expressed in a subset of blood vessels (arrows in a and i) that were occludin-negative. Occludin was concentrated at TJs in distal tubules in the kidney (arrowheads in b) and intestinal epithelial cells (arrowheads in j). In the kidney, VE-cadherin–positive intertubular capillaries (arrowheads in d and f) and glomerular capillaries (asterisk in d and f) did not express claudin-5/TMVCF, but afferent and efferent arterioles (arrows in c and d) expressed both claudin-5/TMVCF and VE-cadherin. When transverse frozen sections of the thicker artery and vein (arrows and arrowheads, respectively, in g and h) of the kidney were stained with anti–claudin-5/6 pAb, only the artery was intensely stained (g). (h) Phase–contrast image. Bars: 40 μm for a–f (f); 70 μm in g and h (h); 40 μm in i and j (j). Cln, claudin; Cad, cadherin.

Mentions: In the kidney, claudin-5/TMVCF did not appear to be expressed in endothelial cells of all segments of the blood vessels (Fig. 5). When frozen sections of the kidney cortex were doubly-stained with anti–claudin-5/6 pAb and antioccludin mAb, some blood vessels running in the connective tissues surrounding renal tubules were stained with anti–claudin-5/6 pAb but not with antioccludin mAb (Fig. 5, a and b). Instead, antioccludin mAb labeled TJs of distal tubules intensely (Fig. 5 b). Anti–claudin-6 pAb showed no significant signals (Fig. 5 e), indicating that only claudin-5/TMVCF was detected in the kidney by the anti–claudin-5/6 pAb. Judging from the density/number of the claudin-5/TMVCF–positive blood vessels, only some selected vessels appeared to be stained with anti–claudin-5/6 pAb. Frozen sections of the kidney were double stained with anti–claudin-5/6 pAb and anti–VE-cadherin mAb (Fig. 5c and Fig. d). Anti–VE-cadherin mAb stained numerous capillaries surrounding renal tubules and in glomeruli. These intertubular and glomerular capillaries were not stained by anti–claudin-5/6 pAb. Interestingly, afferent as well as efferent arterioles of glomeruli were reproducibly stained with anti–claudin-5/6 pAb (Fig. 5c and Fig. d). Furthermore, thicker arteries (probably interlobular arteries) but not veins were also intensely stained with anti–claudin-5/6 pAb (Fig. 5g and Fig. h). Taken together, we concluded that in the kidney, claudin-5/TMVCF constituted TJs only in endothelial cells of the arteries.


Endothelial claudin: claudin-5/TMVCF constitutes tight junction strands in endothelial cells.

Morita K, Sasaki H, Furuse M, Tsukita S - J. Cell Biol. (1999)

Localization of claudin-5/TMVCF in the kidney and the intestine. Frozen sections of the kidney (a–h) and the intestine (i and j) were double labeled with anti–claudin-5/6 pAb (a) and antioccludin mAb (b), anti–claudin-5/6 pAb (c) and anti–VE-cadherin mAb (d), anti–claudin-6 pAb (e) and anti–VE-cadherin mAb (f), or anti–claudin-5/6 pAb (i) and antioccludin mAb (j). Since anti–claudin-6 pAb yielded no signals in the kidney (e) and the intestine (data not shown), the anti–claudin-5/6 pAb staining can be considered to represent the distribution of claudin-5/TMVCF. Claudin-5/TMVCF appeared to be expressed in a subset of blood vessels (arrows in a and i) that were occludin-negative. Occludin was concentrated at TJs in distal tubules in the kidney (arrowheads in b) and intestinal epithelial cells (arrowheads in j). In the kidney, VE-cadherin–positive intertubular capillaries (arrowheads in d and f) and glomerular capillaries (asterisk in d and f) did not express claudin-5/TMVCF, but afferent and efferent arterioles (arrows in c and d) expressed both claudin-5/TMVCF and VE-cadherin. When transverse frozen sections of the thicker artery and vein (arrows and arrowheads, respectively, in g and h) of the kidney were stained with anti–claudin-5/6 pAb, only the artery was intensely stained (g). (h) Phase–contrast image. Bars: 40 μm for a–f (f); 70 μm in g and h (h); 40 μm in i and j (j). Cln, claudin; Cad, cadherin.
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Related In: Results  -  Collection

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Figure 5: Localization of claudin-5/TMVCF in the kidney and the intestine. Frozen sections of the kidney (a–h) and the intestine (i and j) were double labeled with anti–claudin-5/6 pAb (a) and antioccludin mAb (b), anti–claudin-5/6 pAb (c) and anti–VE-cadherin mAb (d), anti–claudin-6 pAb (e) and anti–VE-cadherin mAb (f), or anti–claudin-5/6 pAb (i) and antioccludin mAb (j). Since anti–claudin-6 pAb yielded no signals in the kidney (e) and the intestine (data not shown), the anti–claudin-5/6 pAb staining can be considered to represent the distribution of claudin-5/TMVCF. Claudin-5/TMVCF appeared to be expressed in a subset of blood vessels (arrows in a and i) that were occludin-negative. Occludin was concentrated at TJs in distal tubules in the kidney (arrowheads in b) and intestinal epithelial cells (arrowheads in j). In the kidney, VE-cadherin–positive intertubular capillaries (arrowheads in d and f) and glomerular capillaries (asterisk in d and f) did not express claudin-5/TMVCF, but afferent and efferent arterioles (arrows in c and d) expressed both claudin-5/TMVCF and VE-cadherin. When transverse frozen sections of the thicker artery and vein (arrows and arrowheads, respectively, in g and h) of the kidney were stained with anti–claudin-5/6 pAb, only the artery was intensely stained (g). (h) Phase–contrast image. Bars: 40 μm for a–f (f); 70 μm in g and h (h); 40 μm in i and j (j). Cln, claudin; Cad, cadherin.
Mentions: In the kidney, claudin-5/TMVCF did not appear to be expressed in endothelial cells of all segments of the blood vessels (Fig. 5). When frozen sections of the kidney cortex were doubly-stained with anti–claudin-5/6 pAb and antioccludin mAb, some blood vessels running in the connective tissues surrounding renal tubules were stained with anti–claudin-5/6 pAb but not with antioccludin mAb (Fig. 5, a and b). Instead, antioccludin mAb labeled TJs of distal tubules intensely (Fig. 5 b). Anti–claudin-6 pAb showed no significant signals (Fig. 5 e), indicating that only claudin-5/TMVCF was detected in the kidney by the anti–claudin-5/6 pAb. Judging from the density/number of the claudin-5/TMVCF–positive blood vessels, only some selected vessels appeared to be stained with anti–claudin-5/6 pAb. Frozen sections of the kidney were double stained with anti–claudin-5/6 pAb and anti–VE-cadherin mAb (Fig. 5c and Fig. d). Anti–VE-cadherin mAb stained numerous capillaries surrounding renal tubules and in glomeruli. These intertubular and glomerular capillaries were not stained by anti–claudin-5/6 pAb. Interestingly, afferent as well as efferent arterioles of glomeruli were reproducibly stained with anti–claudin-5/6 pAb (Fig. 5c and Fig. d). Furthermore, thicker arteries (probably interlobular arteries) but not veins were also intensely stained with anti–claudin-5/6 pAb (Fig. 5g and Fig. h). Taken together, we concluded that in the kidney, claudin-5/TMVCF constituted TJs only in endothelial cells of the arteries.

Bottom Line: In contrast, in the kidney, the claudin-5/TMVCF signal was restricted to endothelial cells of arteries, but was undetectable in those of veins and capillaries.Furthermore, when claudin-5/TMVCF cDNA was introduced into mouse L fibroblasts, TJ strands were reconstituted that resembled those in endothelial cells in vivo, i.e., the extracellular face-associated TJs.These findings indicated that claudin-5/TMVCF is an endothelial cell-specific component of TJ strands.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Faculty of Medicine, Kyoto University, Kyoto 606-8501, Japan.

ABSTRACT
Tight junctions (TJs) in endothelial cells are thought to determine vascular permeability. Recently, claudin-1 to -15 were identified as major components of TJ strands. Among these, claudin-5 (also called transmembrane protein deleted in velo-cardio-facial syndrome [TMVCF]) was expressed ubiquitously, even in organs lacking epithelial tissues, suggesting the possible involvement of this claudin species in endothelial TJs. We then obtained a claudin-6-specific polyclonal antibody and a polyclonal antibody that recognized both claudin-5/TMVCF and claudin-6. In the brain and lung, immunofluorescence microscopy with these polyclonal antibodies showed that claudin-5/TMVCF was exclusively concentrated at cell-cell borders of endothelial cells of all segments of blood vessels, but not at those of epithelial cells. Immunoreplica electron microscopy revealed that claudin-5/TMVCF was a component of TJ strands. In contrast, in the kidney, the claudin-5/TMVCF signal was restricted to endothelial cells of arteries, but was undetectable in those of veins and capillaries. In addition, in all other tissues we examined, claudin-5/TMVCF was specifically detected in endothelial cells of some segments of blood vessels, but not in epithelial cells. Furthermore, when claudin-5/TMVCF cDNA was introduced into mouse L fibroblasts, TJ strands were reconstituted that resembled those in endothelial cells in vivo, i.e., the extracellular face-associated TJs. These findings indicated that claudin-5/TMVCF is an endothelial cell-specific component of TJ strands.

Show MeSH
Related in: MedlinePlus