Human Molecular Genetics, 2000, Vol. 9, No. 20 2937-2945
© 2000 Oxford University Press
Mice lacking renal chloride channel, CLC-5, are a model for Dent’s disease, a nephrolithiasis disorder associated with defective receptor-mediated endocytosis
Johns Hopkins University School of Medicine, Departments of Physiology and Medicine, 725 North Wolfe Street, Baltimore, MD 21205, USA, 1Division of Nephrology and Cell Unit, Christian de Duve Institute of Cellular Pathology, Université Catholique de Louvain Medical School, B-1200 Brussels, Belgium and 2Molecular Endocrinology Group, Nuffield Department of Medicine, University of Oxford, John Radcliffe Hospital, Headington, Oxford OX3 9DU, UK
Received 19 October 2000; Revised and Accepted 23 October 2000.
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ABSTRACT |
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Nephrolithiasis (kidney stones) affects 5–10% of adults and is most commonly associated with hypercalciuria, which may be due to monogenic renal tubular disorders. One such hypercalciuric disorder is Dent’s disease, which is characterized by renal proximal tubular defects that include low molecular weight proteinuria, aminoaciduria and glycosuria, together with rickets in some patients. Dent’s disease is due to inactivating mutations of the renal-specific voltage-gated chloride channel, CLC-5, which is expressed in the proximal tubule, thick ascending limb and collecting duct. The subcellular localization of CLC-5 to the proximal tubular endosomes has suggested a role in endocytosis, and to facilitate in vivo investigations of CLC-5 in Dent’s disease we generated mice lacking CLC-5 by targeted gene disruption. CLC-5-deficient mice developed renal tubular defects which included low molecular weight (<70 kDa) proteinuria, generalized aminoaciduria that was more pronounced for neutral and polar amino acids, and glycosuria. They also developed hypercalciuria and renal calcium deposits and some had deformities of the spine. Furthermore, endocytosis as assessed by horseradish peroxidase uptake in the proximal tubule was severely impaired in CLC-5-deficient mice, thereby demonstrating a role for CLC-5 in endosomal uptake of low molecular weight proteins. Thus, CLC-5-deficient mice provide a model for Dent’s disease and this will help in elucidating the function of this chloride channel in endocytosis and renal calcium homeostasis.
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INTRODUCTION |
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Dent’s disease is an X-linked renal tubular disorder characterized by low molecular weight (LMW) proteinuria, aminoaciduria, glycosuria, hypercalciuria, nephrocalcinosis, nephrolithiasis and progressive renal failure, and is complicated by rickets or osteomalacia in some patients (1–3). The aetiology of Dent’s disease, and three other phenotypic variants referred to as X-linked recessive nephrolithiasis (XRN), X-linked recessive hypophosphataemic rickets (XLRH) and the idiopathic LMW proteinuria of Japanese children, has been established to be due to mutations of the CLCN5 gene. CLCN5 encodes CLC-5, which is a member of the CLC family of voltage-gated chloride channels (1,4–7). CLC-5 conducts outwardly rectifying chloride currents that are activated by strong depolarizing voltages and inhibited by DIDS (7). Mutations of CLC-5 that are associated with Dent’s disease abolish or markedly reduce these Cl–currents (1,6,7).
The expression of CLC-5 in the human nephron has been shown to be in the proximal tubule (PT), the thick ascending limb (TAL) and the intercalated cells of the collecting duct (CD) (8). Furthermore, CLC-5 has been localized intracellularly to the PT subapical endosomes and with the vacuolar H+-ATPase, thereby suggesting that it may have a role in the counterion transport mechanism that facilitates acidification within endosomes (8–10). These endosomes form part of the megalin receptor-mediated endocytic pathway, which reabsorbs proteins such as albumin and LMW (<70 kDa) proteins [e.g. 
1-microglobulin, ß2-microglobulin, vitamin D binding protein (DBP) and Clara cell protein (CC16)] that are freely filtered (11). Thus, CLC-5 dysfunction in this pathway may be associated with the observed PT defects such as LMW proteinuria in Dent’s disease. In addition, CLC-5 mRNA expression in the renal cortex is regulated by parathyroid hormone (PTH), thereby also supporting a role for CLC-5 in calcium homeostasis (12), but the mechanisms whereby CLC-5 dysfunction results in hypercalciuria and the other features of Dent’s disease remain to be elucidated. The availability of an animal model for Dent’s disease would greatly facilitate these studies, and we therefore undertook to establish a mouse lacking CLC-5. Mice deficient for CLC-5 were found to show remarkable similarities to Dent’s disease, and demonstrated that CLC-5 is involved in receptor-mediated endocytosis and calcium homeostasis.
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RESULTS |
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Clcn5-deficient mice were generated by targeted deletion of exon VI, which encodes the putative third and fourth transmembrane domains of the CLC-5 channel (1,4) and replacement of these sequences by a neomycin resistance gene in embryonic stem (ES) cells (Fig. 1A). A total of 355 independently targeted ES cells were generated and Southern blot analysis using an external 3' probe and polymerase chain reaction (PCR) analysis using internal primers confirmed the homologous recombination events (Fig. 1B). Four ES clones were microinjected into C57BL/6J mouse blastocysts to produce male chimeras which were mated to C57BL/6J female mice. Germline transmission was obtained from one of the injected ES cell lines and offspring were genotyped (Fig. 1B). All F1 female mice were heterozygotes (+/–) and all male mice were wild-type (+/Y), consistent with an X-linked inheritance. Mating between F1 +/– female and +/Y male mice yielded 249 F2 progeny consisting of: 24.1% +/+; 20.1% +/–; 31.7% +/Y; 19.3% –/Y; and 4.8% deaths. Mating between F2 +/– and –/Y mice yielded 237 F3 progeny consisting of: 25.3% +/–; 16.9% –/–; 24.1% +/Y; 20.7% –/Y; and 13.0% deaths. These distributions are consistent with a Mendelian inheritance. Most of the Clcn5-deficient mice were viable at birth with normal growth and survival to reproductive age (6–9 weeks). Northern (Fig. 1C) and western (Fig. 1D) blot analyses of kidneys revealed an absence of CLC-5 in the –/Y and –/– mice.
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A small number of mutant mice had deformities of the dorsal spine (Fig. 2A) and backward growth of the teeth (data not shown), but radiological evidence for osteopenia or rickets was absent. No gross morphological abnormalities were detected by radiology and magnetic resonance imaging of the kidneys, but histochemistry demonstrated the presence of microscopic calcium deposits at the cortico-medullary junction (Fig. 2B), consistent with the nephrocalcinosis observed in patients with Dent’s disease (3). Renal histology in –/Y mice was normal, but immunostaining with anti-CLC-5 antibodies verified that CLC-5 was absent from the mutant kidney (Fig. 3A). In contrast, the expression and polarity of PT markers including aminopeptidase N (a marker of the apical pole of PT epithelial cells), vacuolar H+-ATPase (a marker of the apical membrane and endosomal compartment) and Na+/K+-ATPase (a marker of the basolateral membrane) were preserved.
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Plasma and urine analyses (Table 1) revealed that the –/Y mice had significant polyuria, hypercalciuria, phosphaturia, proteinuria and glycosuria that was not associated with hyperglycaemia. The polyuria, which correlated with glycosuria (r = 0.4, P < 0.03), resulted in a daily diuresis that was 50% greater than in control littermates and equivalent to 30% of body weight in some –/Y mice. The daily urinary excretion of calcium was
2-fold higher in Clcn5-deficient mice, an increase that is similar to that observed in patients with Dent’s disease (3,13,14). Renal function, assessed by plasma creatinine and creatinine clearance standardized for body weight, remained normal. The –/Y mice also had generalized aminoaciduria (Fig. 4A), and this was more pronounced for neutral and polar amino acids, e.g. citrulline (94-fold increase over +/Y mice matched for age), hydroxyproline (24-fold increase), cysteine (22-fold increase), glutamine (11-fold increase) and threonine (10-fold increase). In contrast, plasma levels of amino acids were similar in +/Y and –/Y mice. Thus, Clcn5-deficient mice develop a phenotype that is consistent with Dent’s disease and have features of the renal Fanconi syndrome which is predominantly a renal PT disorder (15).
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The increased urinary protein excretion observed in –/Y mice (Table 1) was further characterized (Fig. 4B and C). The results showed that the mutant mice had an LMW proteinuria (Fig. 4B) which included DBP (50 kDa) and CC16 (16 kDa). Using CC16 excretion as a measure of LMW proteinuria (Fig. 4C), a marked increase of
200-fold in urinary excretion was observed in the –/Y compared with the +/Y males, and an intermediate increase was observed in the +/– females. Plasma CC16 concentrations were similar in the mutant and wild-type mice (data not shown). These findings, which are consistent with those of an X-linked disorder, are similar to those observed for the excretion of LMW proteins in patients with Dent’s disease and its phenotypic variants, e.g. XRN and the idiopathic LMW proteinuria of Japanese children (5,14,16). LMW proteins are normally filtered at the glomerulus and avidly recaptured by megalin receptor-mediated endocytosis in PT cells (11). CLC-5 has been localized to early endosomes that form part of this pathway (8). In order to study this process more closely, mice were injected with horseradish peroxidase, a classic endocytic tracer of 40 kDa that is readily filtered by the glomerulus and taken up by endocytosis in PT cells (11). Cytochemistry and electron microscopy (Fig. 5) showed a severe impairment of protein endocytosis by PT cells in –/Y mice, such that peroxidase bound to the brush border was poorly transferred into early endocytic vesicles, and escaped the PT segment to become detectable in distal segments of the nephron. Thus, an impairment of receptor-mediated endocytosis in PT cells of Clcn5-deficient mice and patients with Dent’s disease would provide a basis for the defective uptake that would result in the increased urinary excretion of LMW proteins.
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DISCUSSION |
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Our studies have established a mouse model for Dent’s disease. This mouse, which is deficient for CLC-5, develops LMW proteinuria and aminoaciduria (Fig. 4), glycosuria and hypercalciuria (Table 1) and nephrocalcinosis (Fig. 2). Renal failure, which is generally only observed in adults with Dent’s disease (3), was not observed in the CLC-5-deficient mice, and this may be due partly to the young age (5–9 weeks) of the mice at the time of assessment. Many of the manifestations observed in patients with Dent’s disease and the CLC-5-deficient mice are due to renal PT dysfunction. Our CLC-5-deficient mouse model has enabled us to further study the role of CLC-5 in the PT (Fig. 5). CLC-5 is located with the H+-ATPase in the early endosomes that form part of the receptor-mediated endocytic pathway (8,9,11) in PT cells, and these findings have suggested a role for CLC-5 in endosomal acidification. Endosomes vary in their degree of acidification (17), depending on whether their electrogenic H+-ATPase is antagonized by other transporters (e.g. by the electrogenic sodium pump) or facilitated by a coupled chloride conductance (18). Inhibition of endosomal acidification prevents the progression of early endosomes to late endosomes/lysosomes, presumably by impairing recruitment of regulatory or coat proteins (19). It is still unknown whether luminal acidification is required for the formation of primary endocytic vesicles, or if poor entry in the endocytic apparatus is a secondary defect, for example due to inefficient recycling of a rate-limiting partner of the endocytic machinery. At least two choride channels have been implicated in acidification of PT cell endosomes. A cAMP-activated chloride conductance has been demonstrated in endosomes (20) and functional defects in the cystic fibrosis transmembrane conductance regulator (CFTR) have been associated with impaired endosomal acidification (21). However, patients with cystic fibrosis have no renal phenotype and little or no increase in urinary protein excretion (22), suggesting that CFTR does not play a key role in PT endocytosis. Data from several sources (8–10) and also presented here indicate that CLC-5 is presumably a more important alternative. Thus, CLC-5 is abundantly expressed in the subapical cytoplasm of PT cells of mice, rats and man and it colocalizes with endocytic tracers and markers (8–10). Furthermore, CLC-5 accumulates in the enlarged early endosomes of a dominant-positive Rab-5 mutant in transfected cells (9). In addition, we show here that its genetic ablation impairs receptor-mediated endocytosis. All these data indicate that CLC-5 is the key mediator of chloride conductance that is necessary for early endosomal acidification and receptor-mediated reabsorption in PT cells. A failure in this process would not only lead to LMW proteinuria but also a loss of amino acids and glucose, presumably by a failure to recycle their specific transporters to the apical cell membrane (23).
A certain degree of calcium reabsorption in the PT occurs via the megalin receptor-mediated endocytosis (11), and although this may account for part of the observed hypercalciuria, it is important to note that regulated calcium reabsorption occurs in the TAL and distal tubule, and the role of CLC-5 in these segments still remains to be elucidated (12). Interestingly, Clcn5-deficient mice and patients with Dent’s disease (3) maintain a normal plasma calcium despite hypercalciuria, and it has been suggested that this is related to an increased intestinal uptake of calcium (16,24). This increased intestinal calcium uptake may be due in part to the higher observed circulating 1,25-(OH)2-vitamin D3 concentrations in patients with Dent’s disease (16), and the basis of this in the face of an increased loss of DBP (which facilitates the entry of 25-OH-vitamin D3, the substrate for the 1-hydroxylase in PT cells) remains to be defined (25). Our establishment of a mouse model which lacks CLC-5 and develops the phenotypic features of Dent’s disease that include LMW proteinuria and hypercalciuria will help in further elucidating the role of this channel in the regulation of solute reabsorption by the renal tubule.
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MATERIALS AND METHODS |
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TOPABSTRACTINTRODUCTIONRESULTSDISCUSSIONMATERIALS AND METHODSREFERENCES |
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Targeting vector
Genomic clones containing portions of the murine Clcn5 gene were isolated from a Lambda Fix II murine 129/Sv genomic library using a 578 bp mouse cDNA Clcn5 probe generated by RT–PCR utilizing primers 5'-CGA GAC CGA GAT AGG CAC CG-3' and 5'-GCA GCA AGC CAC GTG CAC TA-3'. Two clones were obtained and the 16 kb genomic DNA fragment containing exons IV, V and VI was subcloned into pBluescript SK+ (Stratagene, La Jolla, CA) and restriction mapped (Fig. 1A). To generate the Clcn5-targeting vector, exon VI was disrupted by introduction of a selectable PGK-Neo cassette, in the same transcriptional orientation as Clcn5 into the unique ApaI site at codon 868 and then the disrupted exon VI was inserted in the Eco47III site in intron V. A tk-DTA negative selection cassette was inserted in the Eco47III site in intron IV at the 5' terminus of the long homology of the targeting construct of the Clcn5 genomic clone and the NotI site in the plasmid polylinker (Fig. 1A). The final 16 kb construct was linearized with NotI for transfection.
Cell culture, transfection and selection
R1 ES cells were cultured on mitotically inactivated mouse embryonic fibroblasts (MIMEFs) in standard ES cell medium (22). ES cells were electroporated (250 V, 0.32 kV) with 25 µg of linearized DNA construct and plated on MIMEFs in ES cell medium with murine leukaemia inhibitory factor (1 x 103 U/ml). G418 (250 µg/ml) was added 24 h post-transfection and the cells were maintained under selection for 6 days. Clones were picked at day 7 and grown without selection drugs on MIMEFs.
Southern blot analysis to identify targeted clones
Genomic DNA was isolated from ES cells (26) and digested with SacI, and fragments were separated on a 0.8% agarose gel. The DNA was transferred to Hybond membrane (Amersham, Little Chalfont, UK) and probed with a 600 bp HincII–SacI genomic fragment downstream to the targeting construct (Fig. 1A). Blots were prehybridized and hybridized in Rapid-hyb buffer (Amersham).
Generation of Clcn5-deficient mice
ES cells of G418-resistant, homologous recombinant clones were karyotyped and used for injection into 3.5-day-old blastocysts isolated from pregnant C57BL/6 females. Male chimeric mice were mated with C57BL/6J females (Jackson Laboratory, Bar Harbor, ME) to obtain +/– animals, which were interbred to generate hemizygous and homozygous mutant progeny. Germline transmission was observed in the coat colour of the F1 offspring. Genomic DNA was isolated from tail biopsies and assessed by Southern blot analysis as described above. Genotyping of subsequent generations was performed by PCR using primers (Fig. 1) from intron V (5'-GCT CTT GCT TCT TCA TCG ATA CAC-3') and from exon VI (5'-ACA GCA GCA AGC CAC GTG CAC CA-3'), yielding 350 bp wild-type fragments; and from the PGK-Neo cassette (5'-ATG CGG TGG GCT CTA TGG CTT CTG-3') and from intron VI, outside the homologous region (5'-TGA GTG CAG GCA CAC GTA TGT GCA-3'), yielding 1500 bp mutant fragments. PCR conditions were 35 cycles of 94°C for 1 min, 60°C for 1 min, and 72°C for 2 min.
Northern blot analysis
Total RNA was prepared from whole mouse kidneys by the Trizol protocol (Gibco BRL, Rockville, MD) and separated (20 µg/lane) on a 1% agarose gel containing 2.2 M formaldehyde. After overnight transfer to Nytran filters (Amersham), CLC-5 transcript was detected after 2 h hybridization at 65°C with a 519 bp Clcn5 cDNA fragment (base pairs 1240–1759), which was generated by RT–PCR from kidney RNA.
Immunostaining
Tissue blocks were prepared from adult mouse kidneys (+/Y and –/Y, n = 6 in each group) to perform immunohistochemistry as described (8), using affinity-purified rabbit polyclonal antibodies (Eurogentec, Seraing, Belgium) against the first extracellular loop (8) or the N-terminus of human CLC-5 (O. Devuyst, in preparation); a monoclonal antibody against the vacuolar H+-ATPase (27); a polyclonal antibody against aminopeptidase N (28); and a monoclonal antibody against the 1 subunit of the Na+/K+-ATPase (Upstate Biotechnology, Lake Placid, NY). Efficiency of transfer was assessed by Ponceau red (Sigma, St Louis, MO) and/or reprobing with a monoclonal antibody against ß-actin (Sigma). Negative controls were obtained with control or pre-immune IgG, or pre-adsorbed anti-CLC-5 antibodies. Overall histology of mouse kidneys was evaluated after haemalun–eosine staining, whereas calcium deposits were visualized after Von Kossa staining. Kidney samples from a patient with nephrocalcinosis were used as a control for the Von Kossa staining procedure.
Peroxidase cytochemistry
Mice (+/Y, n = 7; –/Y, n = 9) were anaesthetized under halothane (Sigma) and injected intravenously with 120 µg/g body wt horseradish peroxidase (Roche Diagnostics, Brussels, Belgium). Five minutes after injection, kidneys were excised, fixed for 6 h at 4°C in 4% paraformaldehyde (Boehringer, Heidelberg, Germany) and rinsed in phosphate-buffered saline (PBS). Peroxidase cytochemistry was performed as described (29). For light microscopy, 1 µm thick sections were counterstained with Toluidine blue. Alternatively, 70 nm thick sections were stained with 2% uranyl acetate and Reynold’s lead citrate and examined in a Philips CM12 electron microscope in the transmission mode under 80 kV.
Urine and plasma analyses
Mice were placed in metabolic cages with ad libidum access to normal food (Lab Diet-PMI Nutrition International, Elkridge, MD) and water. Twenty-four hour urine collections were obtained, and blood was obtained by cardiac puncture. Serum and urine levels of CC16, a 16 kDa marker for early detection of tubular dysfunction (30), were determined in duplicate by immunoassay (31). Plasma and urinary electrolytes and creatinine were measured by standard methods (Anilab, Baltimore, MD). Urine was also analysed by 10% SDS–PAGE and immunoblotting as described (8,25), using affinity-purified rabbit antibodies against (i) human LMWP; (ii) human CC16; and (iii) human DBP (Dako, Glostrup, Denmark).
GenBank accession number
The sequence of rat CLC-5 mRNA was deposited in GenBank under accession no. D50497.1.
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ACKNOWLEDGEMENTS |
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The dedicated help of V. Cebotaru, Y. Cnops, X. Dumont, G. Gou, M. Leruth, L. Tanh and P. van Der Smissen is acknowledged. We are indebted to C. Hermans, R. Beauwens, M. Stoenoiu, M.-F. Vincent and C. van Ypersele de Strihou for helpful discussions and suggestions. We thank E.F. McCarthy Jr for radiological investigations, and S. Lee, R. Reeves, S. Gluck and A. Hubbard for the gift of reagents. The blastocyst injections were performed by the Transgenic Mice Core Facility at Johns Hopkins University. This work was funded in part by NIH DK 43423 (S.E.G.); NIH DK32753 (W.B.G.); Fondation Alphonse & Jean Forton, FRSM, FNRS and Concerted Research Action (O.D.); FNRS, Concerted Research Action and Interuniversity Attraction Pole (P.J.C.); Medical Research Council of the UK (R.V.T.).
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FOOTNOTES |
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+ These authors contributed equally to this work
§ To whom correspondence should be addressed. Tel: +1 410 955 7166; Fax: +1 410 955 0461; Email: wguggino@bs.jhmi.edu
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REFERENCES |
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 T. Maritzen, D. J. Keating, I. Neagoe, A. A. Zdebik, and T. J. Jentsch Role of the Vesicular Chloride Transporter ClC-3 in Neuroendocrine Tissue J. Neurosci., October 15, 2008; 28(42): 10587 - 10598. [Abstract] [Full Text] [PDF]
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 J. Wright, M. M. Morales, J. Sousa-Menzes, D. Ornellas, J. Sipes, Y. Cui, I. Cui, P. Hulamm, V. Cebotaru, L. Cebotaru, et al. Transcriptional adaptation to Clcn5 knockout in proximal tubules of mouse kidney Physiol Genomics, May 1, 2008; 33(3): 341 - 354. [Abstract] [Full Text] [PDF]
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 T.-Y. Chen and T.-C. Hwang CLC-0 and CFTR: Chloride Channels Evolved From Transporters Physiol Rev, April 1, 2008; 88(2): 351 - 387. [Abstract] [Full Text] [PDF]
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 S. Terryn, F. Jouret, F. Vandenabeele, I. Smolders, M. Moreels, O. Devuyst, P. Steels, and E. V. Kerkhove A primary culture of mouse proximal tubular cells, established on collagen-coated membranes Am J Physiol Renal Physiol, August 1, 2007; 293(2): F476 - F485. [Abstract] [Full Text] [PDF]
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 R. Nielsen, P. J. Courtoy, C. Jacobsen, G. Dom, W. R. Lima, M. Jadot, T. E. Willnow, O. Devuyst, and E. I. Christensen Endocytosis provides a major alternative pathway for lysosomal biogenesis in kidney proximal tubular cells PNAS, March 27, 2007; 104(13): 5407 - 5412. [Abstract] [Full Text] [PDF]
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 F. Jouret, A. Bernard, C. Hermans, G. Dom, S. Terryn, T. Leal, P. Lebecque, J.-J. Cassiman, B. J. Scholte, H. R. de Jonge, et al. Cystic Fibrosis Is Associated with a Defect in Apical Receptor-Mediated Endocytosis in Mouse and Human Kidney J. Am. Soc. Nephrol., March 1, 2007; 18(3): 707 - 718. [Abstract] [Full Text] [PDF]
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 T. J. Jentsch Chloride and the endosomal-lysosomal pathway: emerging roles of CLC chloride transporters J. Physiol., February 1, 2007; 578(3): 633 - 640. [Abstract] [Full Text] [PDF]
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M. Ludwig, B. Utsch, and L. A. H. Monnens Recent advances in understanding the clinical and genetic heterogeneity of Dent's disease Nephrol. Dial. Transplant., October 1, 2006; 21(10): 2708 - 2717. [Full Text] [PDF]
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M. Poet, U. Kornak, M. Schweizer, A. A. Zdebik, O. Scheel, S. Hoelter, W. Wurst, A. Schmitt, J. C. Fuhrmann, R. Planells-Cases, et al. Lysosomal storage disease upon disruption of the neuronal chloride transport protein ClC-6 PNAS, September 12, 2006; 103(37): 13854 - 13859. [Abstract] [Full Text] [PDF]
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E. Tosetto, G. M. Ghiggeri, F. Emma, G. Barbano, A. Carrea, G. Vezzoli, R. Torregrossa, M. Cara, G. Ripanti, A. Ammenti, et al. Phenotypic and genetic heterogeneity in Dent's disease--the results of an Italian collaborative study Nephrol. Dial. Transplant., September 1, 2006; 21(9): 2452 - 2463. [Abstract] [Full Text] [PDF]
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 C. Sirac, F. Bridoux, C. Carrion, O. Devuyst, B. Fernandez, J.-M. Goujon, C. E. Hamel, J.-C. Aldigier, G. Touchard, and M. Cogne Role of the monoclonal {kappa} chain V domain and reversibility of renal damage in a transgenic model of acquired Fanconi syndrome Blood, July 15, 2006; 108(2): 536 - 543. [Abstract] [Full Text] [PDF]
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 D. H. Hryciw, J. Ekberg, C. Ferguson, A. Lee, D. Wang, R. G. Parton, C. A. Pollock, C. C. Yun, and P. Poronnik Regulation of Albumin Endocytosis by PSD95/Dlg/ZO-1 (PDZ) Scaffolds: INTERACTION OF Na+-H+ EXCHANGE REGULATORY FACTOR-2 WITH ClC-5 J. Biol. Chem., June 9, 2006; 281(23): 16068 - 16077. [Abstract] [Full Text] [PDF]
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 M.-F. van den Hove, K. Croizet-Berger, F. Jouret, S. E. Guggino, W. B. Guggino, O. Devuyst, and P. J. Courtoy The Loss of the Chloride Channel, ClC-5, Delays Apical Iodide Efflux and Induces a Euthyroid Goiter in the Mouse Thyroid Gland Endocrinology, March 1, 2006; 147(3): 1287 - 1296. [Abstract] [Full Text] [PDF]
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N. Picard, M. Van Abel, C. Campone, M. Seiler, M. Bloch-Faure, J. G.J. Hoenderop, J. Loffing, P. Meneton, R. J.M. Bindels, M. Paillard, et al. Tissue Kallikrein-Deficient Mice Display a Defect in Renal Tubular Calcium Absorption J. Am. Soc. Nephrol., December 1, 2005; 16(12): 3602 - 3610. [Abstract] [Full Text] [PDF]
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C. Caruso-Neves, S.-H. Kwon, and W. B. Guggino Albumin endocytosis in proximal tubule cells is modulated by angiotensin II through an AT2 receptor-mediated protein kinase B activation PNAS, November 29, 2005; 102(48): 17513 - 17518. [Abstract] [Full Text] [PDF]
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Y. Wang, H. Cai, L. Cebotaru, D. H. Hryciw, E. J. Weinman, M. Donowitz, S. E. Guggino, and W. B. Guggino ClC-5: role in endocytosis in the proximal tubule Am J Physiol Renal Physiol, October 1, 2005; 289(4): F850 - F862. [Abstract] [Full Text] [PDF]
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T. J. Jentsch Chloride Transport in the Kidney: Lessons from Human Disease and Knockout Mice J. Am. Soc. Nephrol., June 1, 2005; 16(6): 1549 - 1561. [Abstract] [Full Text] [PDF]
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O. W. Moe and O. Bonny Genetic Hypercalciuria J. Am. Soc. Nephrol., March 1, 2005; 16(3): 729 - 745. [Abstract] [Full Text] [PDF]
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D. H. Hryciw, J. Ekberg, A. Lee, I. L. Lensink, S. Kumar, W. B. Guggino, D. I. Cook, C. A. Pollock, and P. Poronnik Nedd4-2 Functionally Interacts with ClC-5: INVOLVEMENT IN CONSTITUTIVE ALBUMIN ENDOCYTOSIS IN PROXIMAL TUBULE CELLS J. Biol. Chem., December 31, 2004; 279(53): 54996 - 55007. [Abstract] [Full Text] [PDF]
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 E. Babini and M. Pusch A Two-Holed Story: Structural Secrets About ClC Proteins Become Unraveled? Physiology, October 1, 2004; 19(5): 293 - 299. [Abstract] [Full Text] [PDF]
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C. A. Wagner, K. E. Finberg, S. Breton, V. Marshansky, D. Brown, and J. P. Geibel Renal Vacuolar H+-ATPase Physiol Rev, October 1, 2004; 84(4): 1263 - 1314. [Abstract] [Full Text] [PDF]
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P.-C. Pham, O. Devuyst, P.-T. Pham, N. Matsumoto, R. N. G. Shih, O. D. Jo, N. Yanagawa, and A. M. Sun Hypertonicity increases CLC-5 expression in mouse medullary thick ascending limb cells Am J Physiol Renal Physiol, October 1, 2004; 287(4): F747 - F752. [Abstract] [Full Text] [PDF]
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P. R. Cutillas, R. J. Chalkley, K. C. Hansen, R. Cramer, A. G. W. Norden, M. D. Waterfield, A. L. Burlingame, and R. J. Unwin The urinary proteome in Fanconi syndrome implies specificity in the reabsorption of proteins by renal proximal tubule cells Am J Physiol Renal Physiol, September 1, 2004; 287(3): F353 - F364. [Abstract] [Full Text] [PDF]
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D. H. Hryciw, Y. Wang, O. Devuyst, C. A. Pollock, P. Poronnik, and W. B. Guggino Cofilin Interacts with ClC-5 and Regulates Albumin Uptake in Proximal Tubule Cell Lines J. Biol. Chem., October 10, 2003; 278(41): 40169 - 40176. [Abstract] [Full Text] [PDF]
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R. Mohammad-Panah, R. Harrison, S. Dhani, C. Ackerley, L.-J. Huan, Y. Wang, and C. E. Bear The Chloride Channel ClC-4 Contributes to Endosomal Acidification and Trafficking J. Biol. Chem., August 1, 2003; 278(31): 29267 - 29277. [Abstract] [Full Text] [PDF]
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E. I. Christensen, O. Devuyst, G. Dom, R. Nielsen, P. Van Der Smissen, P. Verroust, M. Leruth, W. B. Guggino, and P. J. Courtoy Loss of chloride channel ClC-5 impairs endocytosis by defective trafficking of megalin and cubilin in kidney proximal tubules PNAS, July 8, 2003; 100(14): 8472 - 8477. [Abstract] [Full Text] [PDF]
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I. Carballo-Trujillo, V. Garcia-Nieto, F. J. Moya-Angeler, M. Anton-Gamero, C. Loris, S. Mendez-Alvarez, and F. Claverie-Martin Novel truncating mutations in the ClC-5 chloride channel gene in patients with Dent's disease Nephrol. Dial. Transplant., April 1, 2003; 18(4): 717 - 723. [Abstract] [Full Text] [PDF]
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K. Strange From Genes to Integrative Physiology: Ion Channel and Transporter Biology in Caenorhabditis elegans Physiol Rev, April 1, 2003; 83(2): 377 - 415. [Abstract] [Full Text] [PDF]
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K. K. Frick and D. A. Bushinsky Molecular Mechanisms of Primary Hypercalciuria J. Am. Soc. Nephrol., April 1, 2003; 14(4): 1082 - 1095. [Full Text] [PDF]
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 G. Cao, G. Yang, T. L. Timme, T. Saika, L. D. Truong, T. Satoh, A. Goltsov, S. H. Park, T. Men, N. Kusaka, et al. Disruption of the Caveolin-1 Gene Impairs Renal Calcium Reabsorption and Leads to Hypercalciuria and Urolithiasis Am. J. Pathol., April 1, 2003; 162(4): 1241 - 1248. [Abstract] [Full Text] [PDF]
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 C. Isnard-Bagnis, N. Da Silva, V. Beaulieu, A. S. L. Yu, D. Brown, and S. Breton Detection of ClC-3 and ClC-5 in epididymal epithelium: immunofluorescence and RT-PCR after LCM Am J Physiol Cell Physiol, January 1, 2003; 284(1): C220 - C232. [Abstract] [Full Text] [PDF]
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O. Devuyst and W. B. Guggino Chloride channels in the kidney: lessons learned from knockout animals Am J Physiol Renal Physiol, December 1, 2002; 283(6): F1176 - F1191. [Abstract] [Full Text] [PDF]
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F. Thevenod Ion channels in secretory granules of the pancreas and their role in exocytosis and release of secretory proteins Am J Physiol Cell Physiol, September 1, 2002; 283(3): C651 - C672. [Abstract] [Full Text] [PDF]
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T. X. Weng, B. F. Godley, G. F. Jin, N. J. Mangini, B. G. Kennedy, A. S. L. Yu, and N. K. Wills Oxidant and antioxidant modulation of chloride channels expressed in human retinal pigment epithelium Am J Physiol Cell Physiol, September 1, 2002; 283(3): C839 - C849. [Abstract] [Full Text] [PDF]
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T. J. Jentsch, V. Stein, F. Weinreich, and A. A. Zdebik Molecular Structure and Physiological Function of Chloride Channels Physiol Rev, April 1, 2002; 82(2): 503 - 568. [Abstract] [Full Text] [PDF]
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H. Sun, T. Tsunenari, K.-W. Yau, and J. Nathans The vitelliform macular dystrophy protein defines a new family of chloride channels PNAS, March 19, 2002; 99(6): 4008 - 4013. [Abstract] [Full Text] [PDF]
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H. Birn, T. E. Willnow, R. Nielsen, A. G. W. Norden, C. Bonsch, S. K. Moestrup, E. Nexo, and E. I. Christensen Megalin is essential for renal proximal tubule reabsorption and accumulation of transcobalamin-B12 Am J Physiol Renal Physiol, March 1, 2002; 282(3): F408 - F416. [Abstract] [Full Text] [PDF]
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R. Mohammad-Panah, C. Ackerley, J. Rommens, M. Choudhury, Y. Wang, and C. E. Bear The Chloride Channel ClC-4 Co-localizes with Cystic Fibrosis Transmembrane Conductance Regulator and May Mediate Chloride Flux across the Apical Membrane of Intestinal Epithelia J. Biol. Chem., January 4, 2002; 277(1): 566 - 574. [Abstract] [Full Text]
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A. Vandewalle Diversity within the CLC chloride channel family involved in inherited diseases: from plasma membranes to acidic organelles Nephrol. Dial. Transplant., January 1, 2002; 17(1): 1 - 3. [Full Text] [PDF]
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J A Sayer, G S Stewart, S H Boese, M A Gray, S H S Pearce, T H J Goodship, and N L Simmons The voltage-dependent Cl- channel ClC-5 and plasma membrane Cl- conductances of mouse renal collecting duct cells (mIMCD-3) J. Physiol., November 1, 2001; 536(3): 769 - 783. [Abstract] [Full Text] [PDF]
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 H. Fares and I. Greenwald Genetic Analysis of Endocytosis in Caenorhabditis elegans: Coelomocyte Uptake Defective Mutants Genetics, September 1, 2001; 159(1): 133 - 145. [Abstract] [Full Text] [PDF]
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N. K. Wills and P. Fong ClC Chloride Channels in Epithelia: Recent Progress and Remaining Puzzles Physiology, August 1, 2001; 16(4): 161 - 166. [Abstract] [Full Text] [PDF]
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 R. D. Edmonds, I. V. Silva, W. B. Guggino, R. B. Butler, P. L. Zeitlin, and C. J. Blaisdell Pre- and Postnatal Lung Development, Maturation, and Plasticity: ClC-5: ontogeny of an alternative chloride channel in respiratory epithelia Am J Physiol Lung Cell Mol Physiol, March 1, 2002; 282(3): L501 - L507. [Abstract] [Full Text] [PDF]
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