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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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Related in: MedlinePlus

The total number of thymocytes is increased in cKO mice.Total number of thymocytes in tamoxifen-treated F/F;Cre (cKO, n = 6) and littermate F/F controls (Ctrl, n = 6). The number of cells in cKO mice is shown as percent of cells in controls. Data are presented as mean ± SEM. P<0.05. (B) Flow cytometric analysis of thymocyte populations (thymus) and T cell subsets (spleen) in cKO and control (Ctrl) mice. Single cells are displayed on a dot plot of CD4 versus CD8. CD8: (CD8+ single positive); CD4: (CD4+ single positive); DP: (CD4+/CD8+ double positive); DN: (CD4−/CD8− double negative). Experiments were performed three times.
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pone-0020203-g006: The total number of thymocytes is increased in cKO mice.Total number of thymocytes in tamoxifen-treated F/F;Cre (cKO, n = 6) and littermate F/F controls (Ctrl, n = 6). The number of cells in cKO mice is shown as percent of cells in controls. Data are presented as mean ± SEM. P<0.05. (B) Flow cytometric analysis of thymocyte populations (thymus) and T cell subsets (spleen) in cKO and control (Ctrl) mice. Single cells are displayed on a dot plot of CD4 versus CD8. CD8: (CD8+ single positive); CD4: (CD4+ single positive); DP: (CD4+/CD8+ double positive); DN: (CD4−/CD8− double negative). Experiments were performed three times.

Mentions: To investigate possible effects of CAR on thymocytes and leukocytes, littermates of F/F;Cre (cKO, n = 10) and F/F controls (Ctrl, n = 10) were treated with tamoxifen. When the thymus and spleen were removed 2–3 days after the last tamoxifen administration, thymus was found to be notably larger in cKO than in control animals (data not shown). Thymocytes and spleen leukocytes were isolated, counted, and analysed by flow cytometry. The results demonstrated an increase in the total number of thymocytes in cKO compared to control mice (P<0.005) (Fig. 6A). This effect was not seen in +/+;Cre control mice demonstrating that Cre protein alone could not affect the number of thymocytes (98±5.7% of controls). The percentage of cells in the four thymocyte populations was unchanged, indicating that all populations were similarly affected (Fig. 6B). The percentage of different thymocyte populations in cKO vs. controls was as follows. CD8+ single positive: 6.08±1.8 in cKO mice vs. 5.15±1.8 in control mice, CD4+ single positive: 9.63±1.8 vs. 9.70±2.2, CD4+/CD8+ double positive (DP): 73.13±6 vs. 74.85±5.4, CD4−/CD8− double negative (DN): 10.42±3.7 vs. 9.33±1.5. The increased number of thymocytes in cKO mice was not caused by an increase in cell proliferation since flow cytometric analysis did not reveal changes in the percentage of cycling thymocytes in cKO mice as compared to controls (4.5±0.4 vs. 4.2±0.7). In addition, no significant difference was found in the percentage of cycling cells among the four different thymocyte populations (data not shown). The sensitivity of the thymocytes to glucocorticoid-induced apoptosis was similar in cKO and control mice (data not shown). Analysis of spleen leukocytes did not reveal a significant change in the number of T cells (data not shown) or percentage of CD4+ and CD8+ T cells (Fig. 6B). The percentages of different T cell populations in cKO vs. controls were as follows. CD4+: 18±4.1 vs. 16±2.2 and CD8+: 9.5±4 vs. 7.1±2.9. There was also no significant change in the percentage of B cells (47.2±6.1 vs. 46.8±3.7).


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 total number of thymocytes is increased in cKO mice.Total number of thymocytes in tamoxifen-treated F/F;Cre (cKO, n = 6) and littermate F/F controls (Ctrl, n = 6). The number of cells in cKO mice is shown as percent of cells in controls. Data are presented as mean ± SEM. P<0.05. (B) Flow cytometric analysis of thymocyte populations (thymus) and T cell subsets (spleen) in cKO and control (Ctrl) mice. Single cells are displayed on a dot plot of CD4 versus CD8. CD8: (CD8+ single positive); CD4: (CD4+ single positive); DP: (CD4+/CD8+ double positive); DN: (CD4−/CD8− double negative). Experiments were performed three times.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3108585&req=5

pone-0020203-g006: The total number of thymocytes is increased in cKO mice.Total number of thymocytes in tamoxifen-treated F/F;Cre (cKO, n = 6) and littermate F/F controls (Ctrl, n = 6). The number of cells in cKO mice is shown as percent of cells in controls. Data are presented as mean ± SEM. P<0.05. (B) Flow cytometric analysis of thymocyte populations (thymus) and T cell subsets (spleen) in cKO and control (Ctrl) mice. Single cells are displayed on a dot plot of CD4 versus CD8. CD8: (CD8+ single positive); CD4: (CD4+ single positive); DP: (CD4+/CD8+ double positive); DN: (CD4−/CD8− double negative). Experiments were performed three times.
Mentions: To investigate possible effects of CAR on thymocytes and leukocytes, littermates of F/F;Cre (cKO, n = 10) and F/F controls (Ctrl, n = 10) were treated with tamoxifen. When the thymus and spleen were removed 2–3 days after the last tamoxifen administration, thymus was found to be notably larger in cKO than in control animals (data not shown). Thymocytes and spleen leukocytes were isolated, counted, and analysed by flow cytometry. The results demonstrated an increase in the total number of thymocytes in cKO compared to control mice (P<0.005) (Fig. 6A). This effect was not seen in +/+;Cre control mice demonstrating that Cre protein alone could not affect the number of thymocytes (98±5.7% of controls). The percentage of cells in the four thymocyte populations was unchanged, indicating that all populations were similarly affected (Fig. 6B). The percentage of different thymocyte populations in cKO vs. controls was as follows. CD8+ single positive: 6.08±1.8 in cKO mice vs. 5.15±1.8 in control mice, CD4+ single positive: 9.63±1.8 vs. 9.70±2.2, CD4+/CD8+ double positive (DP): 73.13±6 vs. 74.85±5.4, CD4−/CD8− double negative (DN): 10.42±3.7 vs. 9.33±1.5. The increased number of thymocytes in cKO mice was not caused by an increase in cell proliferation since flow cytometric analysis did not reveal changes in the percentage of cycling thymocytes in cKO mice as compared to controls (4.5±0.4 vs. 4.2±0.7). In addition, no significant difference was found in the percentage of cycling cells among the four different thymocyte populations (data not shown). The sensitivity of the thymocytes to glucocorticoid-induced apoptosis was similar in cKO and control mice (data not shown). Analysis of spleen leukocytes did not reveal a significant change in the number of T cells (data not shown) or percentage of CD4+ and CD8+ T cells (Fig. 6B). The percentages of different T cell populations in cKO vs. controls were as follows. CD4+: 18±4.1 vs. 16±2.2 and CD8+: 9.5±4 vs. 7.1±2.9. There was also no significant change in the percentage of B cells (47.2±6.1 vs. 46.8±3.7).

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