Limits...
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.

Show MeSH

Related in: MedlinePlus

Inactivation of the Car gene following tamoxifen administration.(A) Genomic tail DNA was isolated from tamoxifen-treated animals with genotypes F/F (ctrl, lane 1) and F/F;Cre (cKO, lane 2). PCR amplification demonstrated that an efficient excision of exon 2 was specifically obtained in the cKO animals. (B) Western blot analysis of organ extracts from tamoxifen-treated F/F control (Ctrl) and F/F;Cre (cKO) animals. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times. (C) Western blotting of protein-extracts prepared from lungs of tamoxifen-treated mice. No loss of CAR protein is found in animals with genotypes +/+;Cre (lane 1) or F/F (lane 3) as compared to wt controls (lane 2) in the presence of tamoxifen. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3108585&req=5

pone-0020203-g002: Inactivation of the Car gene following tamoxifen administration.(A) Genomic tail DNA was isolated from tamoxifen-treated animals with genotypes F/F (ctrl, lane 1) and F/F;Cre (cKO, lane 2). PCR amplification demonstrated that an efficient excision of exon 2 was specifically obtained in the cKO animals. (B) Western blot analysis of organ extracts from tamoxifen-treated F/F control (Ctrl) and F/F;Cre (cKO) animals. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times. (C) Western blotting of protein-extracts prepared from lungs of tamoxifen-treated mice. No loss of CAR protein is found in animals with genotypes +/+;Cre (lane 1) or F/F (lane 3) as compared to wt controls (lane 2) in the presence of tamoxifen. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times.

Mentions: PCR performed on DNA isolated from the tail demonstrated an efficient excision of the second exon specifically in the cKOs (Fig. 2A). The same result was seen when PCR was performed on DNA isolated from other organs including kidney, heart and spleen (data not shown). One notable exception was the liver, in which the recombination was much less efficient compared to the other organs tested (data not shown). This observation was in accordance with a previous report, demonstrating reduced efficiency of the CMV-Cre-ERTM system in the liver [33]. The low recombination efficiency in the liver might reflect a metabolization and subsequent inactivation of the drug.


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)

Inactivation of the Car gene following tamoxifen administration.(A) Genomic tail DNA was isolated from tamoxifen-treated animals with genotypes F/F (ctrl, lane 1) and F/F;Cre (cKO, lane 2). PCR amplification demonstrated that an efficient excision of exon 2 was specifically obtained in the cKO animals. (B) Western blot analysis of organ extracts from tamoxifen-treated F/F control (Ctrl) and F/F;Cre (cKO) animals. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times. (C) Western blotting of protein-extracts prepared from lungs of tamoxifen-treated mice. No loss of CAR protein is found in animals with genotypes +/+;Cre (lane 1) or F/F (lane 3) as compared to wt controls (lane 2) in the presence of tamoxifen. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0020203-g002: Inactivation of the Car gene following tamoxifen administration.(A) Genomic tail DNA was isolated from tamoxifen-treated animals with genotypes F/F (ctrl, lane 1) and F/F;Cre (cKO, lane 2). PCR amplification demonstrated that an efficient excision of exon 2 was specifically obtained in the cKO animals. (B) Western blot analysis of organ extracts from tamoxifen-treated F/F control (Ctrl) and F/F;Cre (cKO) animals. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times. (C) Western blotting of protein-extracts prepared from lungs of tamoxifen-treated mice. No loss of CAR protein is found in animals with genotypes +/+;Cre (lane 1) or F/F (lane 3) as compared to wt controls (lane 2) in the presence of tamoxifen. The CAR-specific antibody RP291 was used to detect CAR. The same result was obtained with RP1284 (data not shown). Calnexin was used as a loading control. The experiment was performed at least three times.
Mentions: PCR performed on DNA isolated from the tail demonstrated an efficient excision of the second exon specifically in the cKOs (Fig. 2A). The same result was seen when PCR was performed on DNA isolated from other organs including kidney, heart and spleen (data not shown). One notable exception was the liver, in which the recombination was much less efficient compared to the other organs tested (data not shown). This observation was in accordance with a previous report, demonstrating reduced efficiency of the CMV-Cre-ERTM system in the liver [33]. The low recombination efficiency in the liver might reflect a metabolization and subsequent inactivation of the drug.

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