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Disordered calcium crystal handling in antisense CLC-5-treated collecting duct cells

Lookup NU author(s): Professor John Sayer, Dr Georgina Carr, Professor Simon Pearce, Professor Tim Goodship, Professor Nicholas Simmons

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Abstract

Dent's disease, an X-linked tubulopathy secondary to defects in chloride channel CLC-5, is characterised by low molecular weight proteinuria, hypercalciuria, nephrocalcinosis, and renal stones. Mechanisms leading to nephrocalcinosis are unknown. Using a murine collecting duct cell line (mIMCD-3), we confirm endogenous expression of mCLC-5. During transfection of antisense CLC-5, we observe a reduction in CLC-5 protein expression that correlates with a reduction in the number of acidic endosomal compartments, as determined by quantitative analysis of confocal microscope images using LysoTracker Red. Using wheat germ agglutinin-lectin as an endocytic marker, an arrest of endocytosis is observed in antisense CLC-5 treated cells. Exposure of the cell surface to calcium oxalate crystals results in crystal agglomeration in a minority of sense CLC-5 transfectants (45%) and all antisense CLC-5 transfectants. We conclude that expression of CLC-5 in mIMCD-3 cells allows acidification of endosomes and endocytosis, and that disruption of CLC-5 expression causes abnormal crystal agglomeration. © 2002 Elsevier Science (USA). All rights reserved.


Publication metadata

Author(s): Sayer JA, Carr G, Pearce SHS, Goodship THJ, Simmons NL

Publication type: Article

Publication status: Published

Journal: Biochemical and Biophysical Research Communications

Year: 2003

Volume: 300

Issue: 2

Pages: 305-310

ISSN (print): 0006-291X

ISSN (electronic): 1090-2104

Publisher: Academic Press

URL: http://dx.doi.org/10.1016/S0006-291X(02)02837-1

DOI: 10.1016/S0006-291X(02)02837-1

PubMed id: 12504084


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