Abstract
Gitelman syndrome (GS) is an autosomal recessive disorder characterized by hypokalemic metabolic alkalosis in conjunction with significant hypomagnesemia and hypocalciuria. The GS phenotype is caused by mutations in the solute carrier family 12, member 3 (SLC12A3) gene that encodes the thiazide-sensitive NaCl cotransporter (NCC). We analyzed DNA samples of 163 patients with a clinical suspicion of GS by direct sequencing of all 26 exons of the SLC12A3 gene. In total, 114 different mutations were identified, 31 of which have not been reported before. These novel variants include 3 deletions, 18 missense, 6 splice site and 4 nonsense mutations. We selected seven missense mutations to investigate their effect on NCC activity and plasma membrane localization by using the Xenopus laevis oocyte expression system. The Thr392Ile mutant did not display transport activity (probably class 2 mutation), while the Asn442Ser and Gln1030Arg NCC mutants showed decreased plasma membrane localization and consequently function, likely due to impaired trafficking (class 3 mutation). Even though the NaCl uptake was hampered for NCC mutants Glu121Asp, Pro751Leu, Ser475Cys and Tyr489His, the transporters reached the plasma membrane (class 4 mutation), suggesting an effect on NCC regulation or ion affinity. The present study shows the identification of 38 novel mutations in the SLC12A3 gene and provides insight into the mechanisms that regulate NCC.
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Acknowledgements
We are grateful to our colleague H Dimke for technical assistance and helpful suggestions. We thank all the patients, their families and the referring doctors for their participation. This study was financially supported by the Netherlands Organization for Scientific Research (ZonMw 9120.6110; ALW 700.55.302), a European Young Investigator award from the European Science Foundation (JGH), the Dutch Kidney foundation (C03.6017) (RJB), (C08.2252) (PS), The Swiss National Centre of Competence in Research (NCCR) Kidney Control of Homeostasis (Kidney.CH 507236) and the European Union Seventh Framework Programme (FP7/2007–2013; no 246539) (BG) and the European Community, FP7 (EUNEFRON 201590) (NVAMK).
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Glaudemans, B., Yntema, H., San-Cristobal, P. et al. Novel NCC mutants and functional analysis in a new cohort of patients with Gitelman syndrome. Eur J Hum Genet 20, 263–270 (2012). https://doi.org/10.1038/ejhg.2011.189
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DOI: https://doi.org/10.1038/ejhg.2011.189
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