title: Detection of a transcript abnormality in mRNA of the SLC12A3 gene extracted from urinary sediment cells of a patient with Gitelman's syndrome.
abstract: To date, many mutations, including intronic nucleotide changes, in the SLC12A3 gene encoding the thiazide-sensitive sodium-chloride cotransporter (NCCT) have been reported in Gitelman's syndrome (GS) patients. However, it has not been clarified whether intronic nucleotide changes affect mRNA content. Since mRNA analysis is possible only after obtaining renal biopsy specimens, no studies have been conducted to identify transcript abnormalities in GS. In the study reported here, we investigated such transcript abnormalities for the first time by using mRNA expressed in a patient's urinary sediment cells. Direct sequencing analysis of leukocyte DNA disclosed one known missense mutation (R399C) and one known nucleotide change of the splicing acceptor site of intron 13 (1670-1 g > t). mRNA extracted from the urinary sediment cells was analyzed by RT-PCR to determine the pathogenic role of the intron mutation. A fragment encompassing exon 13 to 15 was amplified as two products, one consisting of all three exons and the other lacking only exon 14 in its entirety. Our investigation was the first to demonstrate exon 14 skipping in an NCCT transcript in renal cells. This methodology thus constitutes a potential noninvasive analytical tool for every inherited kidney disease.
authors.affiliation: Department of Pediatrics, Kobe University Graduate School of Medicine, Kobe, Hyogo, Japan.
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Kaito
authors.forename: Kandai
authors.initials: K
authors.lastname: Nozu
authors.forename: Xue J
authors.initials: XJ
authors.lastname: Fu
authors.forename: Ichiro
authors.initials: I
authors.lastname: Kamioka
authors.forename: Teruo
authors.initials: T
authors.lastname: Fujita
authors.forename: Kyoko
authors.initials: K
authors.lastname: Kanda
authors.forename: Rafal P
authors.initials: RP
authors.lastname: Krol
authors.forename: Ryo
authors.initials: R
authors.lastname: Suminaga
authors.forename: Akihito
authors.initials: A
authors.lastname: Ishida
authors.forename: Kazumoto
authors.initials: K
authors.lastname: Iijima
authors.forename: Masafumi
authors.initials: M
authors.lastname: Matsuo
mesh_terms: D000293:Adolescent; D019943:Amino Acid Substitution / Q000235:genetics; D005260:Female; D053579:Gitelman Syndrome / Q000235:genetics* / Q000652:urine; D006801:Humans; D017354:Point Mutation; D012333:RNA, Messenger / Q000235:genetics* / Q000652:urine; D011955:Receptors, Drug / Q000235:genetics*; D064486:Solute Carrier Family 12, Member 3; D027981:Symporters / Q000235:genetics*
chemical_list: D012333:RNA, Messenger; D011955:Receptors, Drug; C578874:SLC12A3 protein, human; D064486:Solute Carrier Family 12, Member 3; D027981:Symporters