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Wnt pathway activation in long term remnant rat model.

Banon-Maneus E, Rovira J, Ramirez-Bajo MJ, Moya-Rull D, Hierro-Garcia N, Takenaka S, Diekmann F, Eickelberg O, Königshoff M, Campistol JM - Biomed Res Int (2014)

Bottom Line: This study aimed to elucidate if the rat 5/6 renal mass reduction model (RMR) is a good model for the Wnt/ β-catenin activation and possible next modulation.Our results demonstrate that Wnt pathway is active by increase of β-catenin at mRNA level and nuclear translocation in tubular epithelium as well as some target genes.These results validate the RMR model for future modulation of Wnt pathway, starting at shorter time after the surgery.

View Article: PubMed Central - PubMed

Affiliation: Comprehensive Pneumology Center, University Hospital Großhadern, Ludwig-Maximilians-University, 81337 Munich, Germany ; Fundació Clínic, Laboratori Experimental de Nefrologia i Transplantament (LENIT), CELLEX 2B, C/Casanova 143, 08036 Barcelona, Spain.

ABSTRACT
Progression of chronic kidney disease (CKD) is characterized by deposition of extracellular matrix. This is an irreversible process that leads to tubulointerstitial fibrosis and finally loss of kidney function. Wnt/ β-catenin pathway was reported to be aberrantly activated in the progressive damage associated with chronic organ failure. Extensive renal ablation is an experimental model widely used to gain insight into the mechanisms responsible for the development of CKD, but it was not evaluated for Wnt/ β-catenin pathway. This study aimed to elucidate if the rat 5/6 renal mass reduction model (RMR) is a good model for the Wnt/ β-catenin activation and possible next modulation. RMR model was evaluated at 12 and 18 weeks after the surgery, when CKD is close to end-stage kidney disease demonstrated by molecular and histological studies. Wnt pathway components were analyzed at mRNA and protein level. Our results demonstrate that Wnt pathway is active by increase of β-catenin at mRNA level and nuclear translocation in tubular epithelium as well as some target genes. These results validate the RMR model for future modulation of Wnt pathway, starting at shorter time after the surgery.

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Related in: MedlinePlus

Activity of the canonical Wnt signal pathway in kidney homogenates of sham and nephrectomized rats. The expression of active Wnt components in kidney homogenates of sham and nephrectomized rats 18 weeks after the surgery was analyzed by immunoblotting of LRP6, phospho-LRP6, total β-catenin, cyclin D1, GSK-3β, and phospho-GSK-3β. Blotting of GDPH and β-actin served as loading controls. Results are derived from 4-5 animals per group.
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fig3: Activity of the canonical Wnt signal pathway in kidney homogenates of sham and nephrectomized rats. The expression of active Wnt components in kidney homogenates of sham and nephrectomized rats 18 weeks after the surgery was analyzed by immunoblotting of LRP6, phospho-LRP6, total β-catenin, cyclin D1, GSK-3β, and phospho-GSK-3β. Blotting of GDPH and β-actin served as loading controls. Results are derived from 4-5 animals per group.

Mentions: To assess Wnt/β-catenin signal activity, Western blot analysis was performed to LRP6, GSK-3β, β-catenin, cyclinD1, and Wnt1 for the 18 weeks' animal groups. Phosphorylation of both LRP6 and GSK-3β was increased (Figure 3), indicating activation of Wnt/β-catenin signaling. This finding correlated with increased expression of total β-catenin and of the Wnt target gene, cyclin D1, both of which were upregulated in Nx rat samples as compared with kidney tissue samples from sham animals. The Wnt ligand Wnt1 at mRNA level showed a tendency to upregulation (log-fold of 2.39 ± 0.87, not statistically significant), while the Wnt1 protein level detected by Western blotting was significantly increased (Figure 4).


Wnt pathway activation in long term remnant rat model.

Banon-Maneus E, Rovira J, Ramirez-Bajo MJ, Moya-Rull D, Hierro-Garcia N, Takenaka S, Diekmann F, Eickelberg O, Königshoff M, Campistol JM - Biomed Res Int (2014)

Activity of the canonical Wnt signal pathway in kidney homogenates of sham and nephrectomized rats. The expression of active Wnt components in kidney homogenates of sham and nephrectomized rats 18 weeks after the surgery was analyzed by immunoblotting of LRP6, phospho-LRP6, total β-catenin, cyclin D1, GSK-3β, and phospho-GSK-3β. Blotting of GDPH and β-actin served as loading controls. Results are derived from 4-5 animals per group.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4066683&req=5

fig3: Activity of the canonical Wnt signal pathway in kidney homogenates of sham and nephrectomized rats. The expression of active Wnt components in kidney homogenates of sham and nephrectomized rats 18 weeks after the surgery was analyzed by immunoblotting of LRP6, phospho-LRP6, total β-catenin, cyclin D1, GSK-3β, and phospho-GSK-3β. Blotting of GDPH and β-actin served as loading controls. Results are derived from 4-5 animals per group.
Mentions: To assess Wnt/β-catenin signal activity, Western blot analysis was performed to LRP6, GSK-3β, β-catenin, cyclinD1, and Wnt1 for the 18 weeks' animal groups. Phosphorylation of both LRP6 and GSK-3β was increased (Figure 3), indicating activation of Wnt/β-catenin signaling. This finding correlated with increased expression of total β-catenin and of the Wnt target gene, cyclin D1, both of which were upregulated in Nx rat samples as compared with kidney tissue samples from sham animals. The Wnt ligand Wnt1 at mRNA level showed a tendency to upregulation (log-fold of 2.39 ± 0.87, not statistically significant), while the Wnt1 protein level detected by Western blotting was significantly increased (Figure 4).

Bottom Line: This study aimed to elucidate if the rat 5/6 renal mass reduction model (RMR) is a good model for the Wnt/ β-catenin activation and possible next modulation.Our results demonstrate that Wnt pathway is active by increase of β-catenin at mRNA level and nuclear translocation in tubular epithelium as well as some target genes.These results validate the RMR model for future modulation of Wnt pathway, starting at shorter time after the surgery.

View Article: PubMed Central - PubMed

Affiliation: Comprehensive Pneumology Center, University Hospital Großhadern, Ludwig-Maximilians-University, 81337 Munich, Germany ; Fundació Clínic, Laboratori Experimental de Nefrologia i Transplantament (LENIT), CELLEX 2B, C/Casanova 143, 08036 Barcelona, Spain.

ABSTRACT
Progression of chronic kidney disease (CKD) is characterized by deposition of extracellular matrix. This is an irreversible process that leads to tubulointerstitial fibrosis and finally loss of kidney function. Wnt/ β-catenin pathway was reported to be aberrantly activated in the progressive damage associated with chronic organ failure. Extensive renal ablation is an experimental model widely used to gain insight into the mechanisms responsible for the development of CKD, but it was not evaluated for Wnt/ β-catenin pathway. This study aimed to elucidate if the rat 5/6 renal mass reduction model (RMR) is a good model for the Wnt/ β-catenin activation and possible next modulation. RMR model was evaluated at 12 and 18 weeks after the surgery, when CKD is close to end-stage kidney disease demonstrated by molecular and histological studies. Wnt pathway components were analyzed at mRNA and protein level. Our results demonstrate that Wnt pathway is active by increase of β-catenin at mRNA level and nuclear translocation in tubular epithelium as well as some target genes. These results validate the RMR model for future modulation of Wnt pathway, starting at shorter time after the surgery.

Show MeSH
Related in: MedlinePlus