Abstract |
Defects in an intracellular chloride channel CLC-5 cause Dent's disease, an inherited kidney stone disorder. Using a collecting duct model, mIMCD-3 cells, we show expression of dimeric mCLC-5. Transient transfection of antisense CLC-5 reduces CLC-5 protein expression. Binding of both calcium phosphate ( hydroxyapatite) and calcium oxalate monohydrate (COM) crystals overlaid onto mIMCD-3 cultures was affected by altered CLC-5 expression. Calcium phosphate crystal agglomerations (>10 microm) were minimal in control (9%) and sense (13%) CLC-5-transfected cells, compared to 66% of antisense CLC-5-transfected cells (P<0.001). Small calcium phosphate crystals (<10 microm) were found associated with 45% of sense CLC-5-treated cells, of which the majority (11/14 cells) appeared to be internalised within the cell. Calcium oxalate agglomerations (>10 microm) were also largely absent for controls or sense mCLC-5 transfectants (11% and 9% of cells, respectively) with COM crystal agglomerates predominating in antisense CLC-5 transfectants (66%, P<0.0001). We conclude that collecting duct cells with reduced CLC-5 expression lead to a tendency to form calcium crystal agglomeration, which may help explain the nephrocalcinosis and nephrolithiasis seen in Dent's disease.
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Authors | J A Sayer, G Carr, N L Simmons |
Journal | Biochimica et biophysica acta
(Biochim Biophys Acta)
Vol. 1689
Issue 1
Pg. 83-90
(May 24 2004)
ISSN: 0006-3002 [Print] Netherlands |
PMID | 15158917
(Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
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Chemical References |
- CLC-5 chloride channel
- Calcium Phosphates
- Chloride Channels
- Calcium Oxalate
- calcium phosphate
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Topics |
- Animals
- Blotting, Western
- Calcium Oxalate
(chemistry, metabolism)
- Calcium Phosphates
(chemistry, metabolism)
- Cell Line
- Chloride Channels
(biosynthesis, genetics, metabolism)
- Crystallization
- Gene Expression Regulation
- Kidney Tubules, Collecting
(cytology, metabolism)
- Mice
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