Functional confirmation of Gitelman's syndrome mutations in Japanese

Gitelman's syndrome is an autosomal recessive inherited renal tubular disorder resulting from loss-of-function mutations in the thiazide-sensitive sodium chloride cotransporter gene (SLC12A3). We have previously reported that the combined allele frequency for the reported Gitelman's syndrome mutations is 0.0321. However, almost all of the reported Gitelman's syndrome mutations were from case reports without functional confirmation. In the present study, we assessed the functionality of the two most prevalent mutations in Japanese, T180K and L849H, using a mammalian cell expression system. Human SLC12A3 cDNA was transiently expressed in Chinese hamster ovary (CHO) cells under the control of a cytomegalo virus (CMV) promoter. The T180K and L849H mutations were introduced by site-directed mutagenesis. The activity of the Na+-Cl- cotransporter was assessed by measuring tracer 22Na+ uptake. While the T180K variation was just a polymorphism, the L849H mutation was confirmed to be a loss-of-function mutation and appears to be responsible for the Gitelman's syndrome. This observation may have very important clinical implications, since the allele frequency of this variation is 0.0126.