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Multiple phenotypes in adult mice following inactivation of the Coxsackievirus and Adenovirus Receptor (Car) gene.

Pazirandeh A, Sultana T, Mirza M, Rozell B, Hultenby K, Wallis K, Vennström B, Davis B, Arner A, Heuchel R, Löhr M, Philipson L, Sollerbrant K - PLoS ONE (2011)

Bottom Line: To determine the normal function of the Coxsackievirus and Adenovirus Receptor (CAR), a protein found in tight junctions and other intercellular complexes, we constructed a mouse line in which the CAR gene could be disrupted at any chosen time point in a broad spectrum of cell types and tissues.All knockouts examined displayed a dilated intestinal tract and atrophy of the exocrine pancreas with appearance of tubular complexes characteristic of acinar-to-ductal metaplasia.These results demonstrate that CAR exerts important functions in the physiology of several organs in vivo.

View Article: PubMed Central - PubMed

Affiliation: Ludwig Institutet for Cancer Research, Stockholm Branch, Stockholm, Sweden.

ABSTRACT
To determine the normal function of the Coxsackievirus and Adenovirus Receptor (CAR), a protein found in tight junctions and other intercellular complexes, we constructed a mouse line in which the CAR gene could be disrupted at any chosen time point in a broad spectrum of cell types and tissues. All knockouts examined displayed a dilated intestinal tract and atrophy of the exocrine pancreas with appearance of tubular complexes characteristic of acinar-to-ductal metaplasia. The mice also exhibited a complete atrio-ventricular block and abnormal thymopoiesis. These results demonstrate that CAR exerts important functions in the physiology of several organs in vivo.

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The integrity of the epithelial lining in intestines of cKO mice is maintained.(A) Tamoxifen-treated F/F;Cre (cKO, n = 11) and littermate F/F controls (Ctrl, n = 11) were given a fluorescent tracer by gavage and the permeability of the intestinal epithelium was analyzed by measuring the presence of fluorescence tracer in serum 4 hrs later. No statistically significant difference was found between the two groups. Data is shown as mean ± SD. P = 0.274. (B) Histology of small intestine harvested two weeks after the last tamoxifen administration from F/F control (Ctrl) and F/F;Cre (cKO) animals. No difference between cKO and control animals was found. Scale bar = 100 µm (C) Indirect immunofluorescence of large intestine from F/F;Cre (cKO) and littermate F/F control (Ctrl) mice two weeks after the last tamoxifen administration. Primary antibodies were directed against CAR (green) and occludin (red). DAPI was used to visualize nuclei (blue). Magnification was 40×. (i) and (ii) are enlargements of the boxed areas in the merged ctrl and cKO images, respectively. The experiment was performed three times.
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pone-0020203-g007: The integrity of the epithelial lining in intestines of cKO mice is maintained.(A) Tamoxifen-treated F/F;Cre (cKO, n = 11) and littermate F/F controls (Ctrl, n = 11) were given a fluorescent tracer by gavage and the permeability of the intestinal epithelium was analyzed by measuring the presence of fluorescence tracer in serum 4 hrs later. No statistically significant difference was found between the two groups. Data is shown as mean ± SD. P = 0.274. (B) Histology of small intestine harvested two weeks after the last tamoxifen administration from F/F control (Ctrl) and F/F;Cre (cKO) animals. No difference between cKO and control animals was found. Scale bar = 100 µm (C) Indirect immunofluorescence of large intestine from F/F;Cre (cKO) and littermate F/F control (Ctrl) mice two weeks after the last tamoxifen administration. Primary antibodies were directed against CAR (green) and occludin (red). DAPI was used to visualize nuclei (blue). Magnification was 40×. (i) and (ii) are enlargements of the boxed areas in the merged ctrl and cKO images, respectively. The experiment was performed three times.

Mentions: The CAR protein is found in the tight junctions of most epithelial cells in vivo and in vitro. A decrease in the amount of CAR protein lowers transepithelial resistance and promotes diffusion of fluorescence markers across epithelial cell layers in vitro [4], [26]. To analyze if inactivation of the Car gene affects epithelial integrity also in vivo, the intestinal epithelium of tamoxifen-treated F/F;Cre (cKO, n = 11) and F/F control mice (Ctrl, n = 11) were challenged with fluorescent tracers administered by gavage. 4 hrs later, blood samples were taken and analyzed for the presence of fluorescent tracer. No statistically significant difference between cKO and control mice was observed indicating a maintained integrity of the intestinal epithelium (Fig. 7A). Similar results were obtained in animals devoid of CAR for up to 18 months (length of experiment). In addition, the cKO mice did not show signs of dehydration or oedema indicating that the skin barrier as well as the vascular system was intact.


Multiple phenotypes in adult mice following inactivation of the Coxsackievirus and Adenovirus Receptor (Car) gene.

Pazirandeh A, Sultana T, Mirza M, Rozell B, Hultenby K, Wallis K, Vennström B, Davis B, Arner A, Heuchel R, Löhr M, Philipson L, Sollerbrant K - PLoS ONE (2011)

The integrity of the epithelial lining in intestines of cKO mice is maintained.(A) Tamoxifen-treated F/F;Cre (cKO, n = 11) and littermate F/F controls (Ctrl, n = 11) were given a fluorescent tracer by gavage and the permeability of the intestinal epithelium was analyzed by measuring the presence of fluorescence tracer in serum 4 hrs later. No statistically significant difference was found between the two groups. Data is shown as mean ± SD. P = 0.274. (B) Histology of small intestine harvested two weeks after the last tamoxifen administration from F/F control (Ctrl) and F/F;Cre (cKO) animals. No difference between cKO and control animals was found. Scale bar = 100 µm (C) Indirect immunofluorescence of large intestine from F/F;Cre (cKO) and littermate F/F control (Ctrl) mice two weeks after the last tamoxifen administration. Primary antibodies were directed against CAR (green) and occludin (red). DAPI was used to visualize nuclei (blue). Magnification was 40×. (i) and (ii) are enlargements of the boxed areas in the merged ctrl and cKO images, respectively. The experiment was performed three times.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3108585&req=5

pone-0020203-g007: The integrity of the epithelial lining in intestines of cKO mice is maintained.(A) Tamoxifen-treated F/F;Cre (cKO, n = 11) and littermate F/F controls (Ctrl, n = 11) were given a fluorescent tracer by gavage and the permeability of the intestinal epithelium was analyzed by measuring the presence of fluorescence tracer in serum 4 hrs later. No statistically significant difference was found between the two groups. Data is shown as mean ± SD. P = 0.274. (B) Histology of small intestine harvested two weeks after the last tamoxifen administration from F/F control (Ctrl) and F/F;Cre (cKO) animals. No difference between cKO and control animals was found. Scale bar = 100 µm (C) Indirect immunofluorescence of large intestine from F/F;Cre (cKO) and littermate F/F control (Ctrl) mice two weeks after the last tamoxifen administration. Primary antibodies were directed against CAR (green) and occludin (red). DAPI was used to visualize nuclei (blue). Magnification was 40×. (i) and (ii) are enlargements of the boxed areas in the merged ctrl and cKO images, respectively. The experiment was performed three times.
Mentions: The CAR protein is found in the tight junctions of most epithelial cells in vivo and in vitro. A decrease in the amount of CAR protein lowers transepithelial resistance and promotes diffusion of fluorescence markers across epithelial cell layers in vitro [4], [26]. To analyze if inactivation of the Car gene affects epithelial integrity also in vivo, the intestinal epithelium of tamoxifen-treated F/F;Cre (cKO, n = 11) and F/F control mice (Ctrl, n = 11) were challenged with fluorescent tracers administered by gavage. 4 hrs later, blood samples were taken and analyzed for the presence of fluorescent tracer. No statistically significant difference between cKO and control mice was observed indicating a maintained integrity of the intestinal epithelium (Fig. 7A). Similar results were obtained in animals devoid of CAR for up to 18 months (length of experiment). In addition, the cKO mice did not show signs of dehydration or oedema indicating that the skin barrier as well as the vascular system was intact.

Bottom Line: To determine the normal function of the Coxsackievirus and Adenovirus Receptor (CAR), a protein found in tight junctions and other intercellular complexes, we constructed a mouse line in which the CAR gene could be disrupted at any chosen time point in a broad spectrum of cell types and tissues.All knockouts examined displayed a dilated intestinal tract and atrophy of the exocrine pancreas with appearance of tubular complexes characteristic of acinar-to-ductal metaplasia.These results demonstrate that CAR exerts important functions in the physiology of several organs in vivo.

View Article: PubMed Central - PubMed

Affiliation: Ludwig Institutet for Cancer Research, Stockholm Branch, Stockholm, Sweden.

ABSTRACT
To determine the normal function of the Coxsackievirus and Adenovirus Receptor (CAR), a protein found in tight junctions and other intercellular complexes, we constructed a mouse line in which the CAR gene could be disrupted at any chosen time point in a broad spectrum of cell types and tissues. All knockouts examined displayed a dilated intestinal tract and atrophy of the exocrine pancreas with appearance of tubular complexes characteristic of acinar-to-ductal metaplasia. The mice also exhibited a complete atrio-ventricular block and abnormal thymopoiesis. These results demonstrate that CAR exerts important functions in the physiology of several organs in vivo.

Show MeSH
Related in: MedlinePlus