J Am Soc Nephrol. 2003 Sep;14(9):2280-7.
Analysis of the polycystins in aortic vascular smooth muscle cells.
Qian, Q., Li, M., Cai, Y., Ward, C. J., Somlo, S., Harris, P. C., Torres, V. E.,
["Division of Nephrology, Mayo Clinic, Rochester, Minnesota 55905, USA. Qian.qi@mayo.edu"]
["Division of Nephrology, Mayo Clinic, Rochester, Minnesota 55905, USA. Qian.qi@mayo.edu"]
The leading cause of death in autosomal dominant polycystic kidney disease (ADPKD) is cardiovascular. However, little is known about the pathogenesis of these manifestations. The present study was undertaken to characterize the ADPKD proteins, the polycystins, in vascular smooth muscle cells. It was demonstrated that the expression of polycystin-1 is developmentally regulated, whereas polycystin-2 has a more constant level of expression. A polycystin-1 subpopulation was immunoprecipitated by polycystin-2, indicating an in vivo interaction of these two proteins. Analysis with glycosidase and cell surface biotinylation indicates that some polycystin-1 products, but not polycystin-2, are located on the plasma membrane. Immunofluorescence showed that most of the polycystin-1 and polycystin-2 was cytoplasmic but that persistent polycystin-1 staining was located in proximity to the cell surface after a Triton-X extraction, whereas no clear surface localization of polycystin-2 was detected. Immuno-gold electron microscopy revealed that polycystin-1 was localized at the plasma membrane and sarcoplasmic reticulum, whereas polycystin-2 was mainly located in the sarcoplasmic reticulum. Both polycystins were found to be associated with dense plaques. These observations are consistent with an important role of the polycystins in the development, maintenance, and function of the myoelastic arterial organization and with the vascular phenotype associated with ADPKD.
PMID: 12937304
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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Validation: In vivo validation
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| Assay with endogenous proteins | Assay with overexpressed proteins | Reference | ||||||||
| Cell or tissue | Cell or tissue | TRP channel construct | Interactor construct | |||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPP1 |
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PKD1/Polycystin-1 | Co-immunofluorescence staining | Mouse vascular smooth muscle cell | 12937304 | |||||
| TRPP1 |
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PKD1/Polycystin-1 | Co-immunoprecipitation | Mouse vascular smooth muscle cell | 12937304 | |||||
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