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Nonspecific cation current associated with native polycystin-2 in HEK-293 cells
Bruna Pelucchi1, Gianluca Aguiari, Angela Pignatelli
1Department of Biology, University of Ferrara, Via L. Borsari 46, Ferrara I-44100, Italy. plb@dns.unife.it
Journal of the American Society of Nephrology : JASN
|January 7, 2006
Summary
Researchers identified a novel cation current (I(CC)) in HEK-293 cells linked to polycystin-2 (PC2) channel activity. This finding offers a new tool for studying autosomal dominant polycystic kidney disease.
Area of Science:
- Molecular Biology
- Cell Physiology
- Genetics
Background:
- Autosomal dominant polycystic kidney disease (ADPKD) is the most common inherited kidney disorder, linked to mutations in PKD1 or PKD2 genes.
- Polycystin-2 (PC2) is a protein encoded by the PKD2 gene and functions as a cation channel.
Purpose of the Study:
- To characterize a nonspecific cation current (I(CC)) in native HEK-293 cells.
- To investigate the association of this current with PC2 channel activity.
- To establish HEK-293 cells as a model for studying ADPKD pathogenesis.
Main Methods:
- Electrophysiological recordings to measure cation current (I(CC)).
- RNA interference (RNAi) to silence PC2 expression.
- Transfection with normal and truncated PC2 constructs.
- Perfusion with antibodies and specific inhibitors.
Main Results:
- A voltage-dependent, inactivating cation current (I(CC)) was identified in native HEK-293 cells.
- PC2 silencing reduced I(CC), which was restored by normal PC2 transfection.
- I(CC) was sensitive to various ion channel blockers (Cd2+, Gd3+, La3+, SKF96365, amiloride).
- Expression of the PC1 cytoplasmic tail enhanced I(CC) amplitude.
Conclusions:
- The identified I(CC) is associated with PC2 channel activity in HEK-293 cells.
- HEK-293 cells provide a valuable model for studying PC2/PC1 complex interactions.
- This research contributes to understanding the molecular mechanisms underlying ADPKD.
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