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Different changes in the expression of multiple kinds of tight-junction proteins during ischemia-reperfusion injury of the rat ileum.

Inoue K, Oyamada M, Mitsufuji S, Okanoue T, Takamatsu T - Acta Histochem Cytochem (2006)

Bottom Line: Dysfunction of tight junctions (TJs), located at the most apical part of the intestinal epithelium, is believed to result in various complications in intestinal disease.From 6 to 24 hours post-reperfusion, villous epithelium was restored by cell migration, and barrier function together with reticular pattern expression of ZO-1, occludin, and claudin-1, -3, and -5, recovered time-dependently.These data demonstrated that recovery of intestinal barrier function is associated with expression of ZO-1, occludin, and claudin-1, -3, and -5, whereas claudin-2 and claudin-4 show unique changes in expression and localization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Cell Regulation, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan.

ABSTRACT
Dysfunction of tight junctions (TJs), located at the most apical part of the intestinal epithelium, is believed to result in various complications in intestinal disease. However, the behaviors of multiple kinds of TJ proteins during ischemia-reperfusion injury are not understood in detail. To determine changes in expression and localization of TJ proteins during intestinal-barrier recovery, we induced intestinal ischemia-reperfusion injury in rats, measured mucosa-to-blood permeability of fluorescein isothiocyanate-dextran-4 kDa, and compared it with spatiotemporal changes of ZO-1, occludin, and claudin-1, -2, -3, -4, and -5 by immunoconfocal microscopy. At 1 hour post-reperfusion, villi were denuded and intestinal-barrier function was lost. From 6 to 24 hours post-reperfusion, villous epithelium was restored by cell migration, and barrier function together with reticular pattern expression of ZO-1, occludin, and claudin-1, -3, and -5, recovered time-dependently. To the contrary, after ischemia-reperfusion injury, the localized expression of claudin-2 and claudin-4 observed in the non-treated control was lost and replaced with broader expression from crypts to villi with increased basolateral claudin-4 expression in epithelial cells. These data demonstrated that recovery of intestinal barrier function is associated with expression of ZO-1, occludin, and claudin-1, -3, and -5, whereas claudin-2 and claudin-4 show unique changes in expression and localization.

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Morphometry of claudin-4 expression. (A) Total gray density value of claudin-4 immunolabeling per cell on villi. * p<0.05 compared with control and 24 hr post-reperfusion. † p<0.05 compared with control and 6 hr post-reperfusion. (B) The number of claudin-4-positive pixels per cell on villi in the binary image. ‡ p<0.05 compared with control. (C) Total gray density value of claudin-4 immunolabeling per area of crypt. (D) The number of claudin-4-positive pixels per area of crypt in the binary image. (C, D) * p<0.05 compared with control and 6 and 24 hr post-reperfusion. † p<0.05 compared with control, 1 hr, and 24 hr post-reperfusion. ‡ p<0.05 compared with 1 hr and 6 hr post-reperfusion. # p<0.05 compared with control and 24 hr post-reperfusion. Scheffé’s test.
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Figure 6: Morphometry of claudin-4 expression. (A) Total gray density value of claudin-4 immunolabeling per cell on villi. * p<0.05 compared with control and 24 hr post-reperfusion. † p<0.05 compared with control and 6 hr post-reperfusion. (B) The number of claudin-4-positive pixels per cell on villi in the binary image. ‡ p<0.05 compared with control. (C) Total gray density value of claudin-4 immunolabeling per area of crypt. (D) The number of claudin-4-positive pixels per area of crypt in the binary image. (C, D) * p<0.05 compared with control and 6 and 24 hr post-reperfusion. † p<0.05 compared with control, 1 hr, and 24 hr post-reperfusion. ‡ p<0.05 compared with 1 hr and 6 hr post-reperfusion. # p<0.05 compared with control and 24 hr post-reperfusion. Scheffé’s test.

Mentions: Because the changes in claudin-4 expression were significantly different from those in other tight-junction proteins, morphometric analysis of claudin-4 expression was performed. In the villi, the total gray density value per cell showed time-dependent increase until 24 hr post reperfusion (Fig. 6A). The increase of the total gray density value of claudin-4 fluorescence per cell indicates that claudin-4 expression level was increased time-dependently during the recovery of epithelium. The number of positive pixels per cell was increased at 6 hr post-reperfusion and kept at the increased level at 24 hr post-reperfusion, demonstrating that the area with claudin-4 expression in the cell was increased during the recovery of epithelium.


Different changes in the expression of multiple kinds of tight-junction proteins during ischemia-reperfusion injury of the rat ileum.

Inoue K, Oyamada M, Mitsufuji S, Okanoue T, Takamatsu T - Acta Histochem Cytochem (2006)

Morphometry of claudin-4 expression. (A) Total gray density value of claudin-4 immunolabeling per cell on villi. * p<0.05 compared with control and 24 hr post-reperfusion. † p<0.05 compared with control and 6 hr post-reperfusion. (B) The number of claudin-4-positive pixels per cell on villi in the binary image. ‡ p<0.05 compared with control. (C) Total gray density value of claudin-4 immunolabeling per area of crypt. (D) The number of claudin-4-positive pixels per area of crypt in the binary image. (C, D) * p<0.05 compared with control and 6 and 24 hr post-reperfusion. † p<0.05 compared with control, 1 hr, and 24 hr post-reperfusion. ‡ p<0.05 compared with 1 hr and 6 hr post-reperfusion. # p<0.05 compared with control and 24 hr post-reperfusion. Scheffé’s test.
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Related In: Results  -  Collection

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Figure 6: Morphometry of claudin-4 expression. (A) Total gray density value of claudin-4 immunolabeling per cell on villi. * p<0.05 compared with control and 24 hr post-reperfusion. † p<0.05 compared with control and 6 hr post-reperfusion. (B) The number of claudin-4-positive pixels per cell on villi in the binary image. ‡ p<0.05 compared with control. (C) Total gray density value of claudin-4 immunolabeling per area of crypt. (D) The number of claudin-4-positive pixels per area of crypt in the binary image. (C, D) * p<0.05 compared with control and 6 and 24 hr post-reperfusion. † p<0.05 compared with control, 1 hr, and 24 hr post-reperfusion. ‡ p<0.05 compared with 1 hr and 6 hr post-reperfusion. # p<0.05 compared with control and 24 hr post-reperfusion. Scheffé’s test.
Mentions: Because the changes in claudin-4 expression were significantly different from those in other tight-junction proteins, morphometric analysis of claudin-4 expression was performed. In the villi, the total gray density value per cell showed time-dependent increase until 24 hr post reperfusion (Fig. 6A). The increase of the total gray density value of claudin-4 fluorescence per cell indicates that claudin-4 expression level was increased time-dependently during the recovery of epithelium. The number of positive pixels per cell was increased at 6 hr post-reperfusion and kept at the increased level at 24 hr post-reperfusion, demonstrating that the area with claudin-4 expression in the cell was increased during the recovery of epithelium.

Bottom Line: Dysfunction of tight junctions (TJs), located at the most apical part of the intestinal epithelium, is believed to result in various complications in intestinal disease.From 6 to 24 hours post-reperfusion, villous epithelium was restored by cell migration, and barrier function together with reticular pattern expression of ZO-1, occludin, and claudin-1, -3, and -5, recovered time-dependently.These data demonstrated that recovery of intestinal barrier function is associated with expression of ZO-1, occludin, and claudin-1, -3, and -5, whereas claudin-2 and claudin-4 show unique changes in expression and localization.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Cell Regulation, Kyoto Prefectural University of Medicine, Kamigyo-ku, Kyoto 602-8566, Japan.

ABSTRACT
Dysfunction of tight junctions (TJs), located at the most apical part of the intestinal epithelium, is believed to result in various complications in intestinal disease. However, the behaviors of multiple kinds of TJ proteins during ischemia-reperfusion injury are not understood in detail. To determine changes in expression and localization of TJ proteins during intestinal-barrier recovery, we induced intestinal ischemia-reperfusion injury in rats, measured mucosa-to-blood permeability of fluorescein isothiocyanate-dextran-4 kDa, and compared it with spatiotemporal changes of ZO-1, occludin, and claudin-1, -2, -3, -4, and -5 by immunoconfocal microscopy. At 1 hour post-reperfusion, villi were denuded and intestinal-barrier function was lost. From 6 to 24 hours post-reperfusion, villous epithelium was restored by cell migration, and barrier function together with reticular pattern expression of ZO-1, occludin, and claudin-1, -3, and -5, recovered time-dependently. To the contrary, after ischemia-reperfusion injury, the localized expression of claudin-2 and claudin-4 observed in the non-treated control was lost and replaced with broader expression from crypts to villi with increased basolateral claudin-4 expression in epithelial cells. These data demonstrated that recovery of intestinal barrier function is associated with expression of ZO-1, occludin, and claudin-1, -3, and -5, whereas claudin-2 and claudin-4 show unique changes in expression and localization.

No MeSH data available.


Related in: MedlinePlus