Expression of apical junction complex proteins in colorectal mucosa of miniature dachshunds with inflammatory colorectal polyps - PubMed (original) (raw)

. 2017 Mar 18;79(3):456-463.

doi: 10.1292/jvms.16-0479. Epub 2017 Jan 16.

Hiroshi Ohta, Yumiko Kagawa, Rommaneeya Leela-Arporn, Angkhana Dermlim, Khoirun Nisa, Tomoya Morita, Tatsuyuki Osuga, Noboru Sasaki, Keitaro Morishita, Kensuke Nakamura, Mitsuyoshi Takiguchi

Affiliations

Expression of apical junction complex proteins in colorectal mucosa of miniature dachshunds with inflammatory colorectal polyps

Nozomu Yokoyama et al. J Vet Med Sci. 2017.

Abstract

We examine the expression of tight junction and adherence junction proteins in the colorectal mucosa of miniature dachshunds (MDs) with inflammatory colorectal polyps (ICRPs). Colorectal mucosa samples were endoscopically obtained from 8 MDs with ICRPs and 8 control dogs for immunoblotting. Paraffin-embedded tissues of surgically resected inflamed lesions from another 5 MDs with ICRPs and full-thickness colorectal specimens from 5 healthy beagles were obtained for immunohistochemistry. The expression patterns of claudin-1, -2, -3, -4, -5, -7 and -8, E-cadherin and β-catenin were analyzed in the non-inflamed mucosa and inflamed mucosa of ICRPs and colorectal mucosa of control dogs by immunoblotting. The localization of these proteins in the inflamed lesions was analyzed by immunohistochemistry. The expressions of each of claudin, E-cadherin and β-catenin were not significantly different between control dogs and non-inflamed colonic mucosa from MDs with ICRPs. In contrast, only E-cadherin and β-catenin were detected in the inflamed lesions of MDs with ICRPs. By immunohistochemistry, claudin-2, -3, -4, -5 and -7, E-cadherin and β-catenin were expressed in the colorectal epithelium within the inflamed mucosa, but not in granulation tissue. Distributions of claudin-2, -3, -4, -5, and -7, E-cadherin and β-catenin in the colonic epithelium were not different between MDs with ICRPs and control dogs. These results indicated that no significant alteration was detected in several tight junction or adherence junction proteins expression in the colorectal epithelium of ICRPs.

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Figures

Fig. 1.

Fig. 1.

Representative photomicrograph of the histologic section of an inflamed lesion of miniature dachshund (MD) with inflammatory colorectal polyps (ICRPs). Histopathological appearance of stage I (A), stage II (B) and stage III (C). (A) Stage I, thickened mucosa composed of hyperplastic goblet cells. (B) Stage II, granulation tissue formation in the apical region of the mucosa and dilated crypts containing abundant mucus. (C) Stage III, excessive production of granulation tissue, abundant neovascularization and marked inflammatory changes. H&E stain; Bar=200 _µ_m for A and 300_µ_m for B and C.

Fig. 2.

Fig. 2.

Photographs of immunoblots that indicate expressions of various tight junction and adherence junction proteins in colorectal mucosa biopsy samples obtained from control dogs, and non-inflamed mucosa and inflamed mucosa of MDs with ICRPs. The photograph of control dogs and non-inflamed mucosa of MDs with ICRPs shows the representative immunoblots of each group. The photograph of inflamed mucosa of MDs with ICRPs shows the results of all of the 8 dogs. β-actin is included as a loading control. Values on the right side indicate the apparent molecular mass (in kilodaltons) of each protein.

Fig. 3.

Fig. 3.

Scatterplots of the expressions of claudin-2, -3, -4, -5 and -7 and E-cadherin; β-catenin in colorectal mucosa samples of control dogs (n=8) and non-inflamed mucosa of MDs with ICRPs (n=8). The horizontal line for each group represents the median. Expression units for each protein relative to β-actin expression.

Fig. 4.

Fig. 4.

Photomicrographs of representative immunostaining results of claudin-2 (A and B), -3 (C and D), -4 (E and F), -5 (G and H) and -7 (I and J) and E-cadherin (K and L); β-catenin (M and N) on the luminal surface of colonic mucosa from control dogs (left column) and stage I to stage II inflamed mucosa (right column) of ICRPs. Bars=50 _µ_m.

Fig. 5.

Fig. 5.

Photomicrographs of representative immunostaining results of claudin-2 (A and B), -3 (C and D), -4 (E and F), -5 (G and H) and -7 (I and J), and E-cadherin (K and L); β-catenin (M and N) in the crypt base of colonic mucosa from control dogs (left column) and stage I to stage II inflamed mucosa (right column) of ICRPs. Bars=50 _µ_m.

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