title: A deletion mutation in Slc12a6 is associated with neuromuscular disease in gaxp mice.
abstract: Giant axonopathy (gaxp), an autosomal recessive mouse mutation, exhibits ataxia of the hind legs with a slight side-to-side wobble while walking. Within the genomic region of the gaxp locus, a total of 94 transcripts were identified; the annotation of these genes using OMIM and PubMed yielded three potential candidate genes. By cDNA microarray analysis, 54 genes located on or near the gaxp locus were found to exhibit differential expression between gaxp and littermate controls. Based on microarray data and the known function of genes identified, Slc12a6 was selected as the primary candidate gene and analyzed using the Reveal technology of SpectruMedix. A 17-base deletion was detected from within exon 4 of Slc12a6. Reverse transcriptase polymerase chain reaction validated the difference in Slc12a6 expression in different types of mice at the mRNA level, revealing a marked reduction in gaxp mice. Western blot analysis indicated that the protein product of Slc12a6, the K(+)-Cl(-) cotransporter Kcc3, was not detectable in gaxp mice. The causative role of the exon 4 mutation within Slc12a6 in the gaxp phenotype was further confirmed by screening multiple inbred strains and by excluding the mutation of nearby genes within the gaxp locus.
authors.affiliation: Department of Orthopaedic Surgery (Campbell Clinic), University of Tennessee Health Science Center, Memphis, TN 38163, USA.
authors.forename: Yan
authors.initials: Y
authors.lastname: Jiao
authors.forename: Xiudong
authors.initials: X
authors.lastname: Jin
authors.forename: Jian
authors.initials: J
authors.lastname: Yan
authors.forename: Chi
authors.initials: C
authors.lastname: Zhang
authors.forename: Feng
authors.initials: F
authors.lastname: Jiao
authors.forename: Xinmin
authors.initials: X
authors.lastname: Li
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Roe
authors.forename: David B
authors.initials: DB
authors.lastname: Mount
authors.forename: Weikuan
authors.initials: W
authors.lastname: Gu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001259:Ataxia / Q000235:genetics*; D001369:Axons / Q000378:metabolism; D004252:DNA Mutational Analysis; D005091:Exons; D017353:Gene Deletion; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D027981:Symporters / Q000235:genetics*; D000096922:K Cl- Cotransporters
chemical_list: D027981:Symporters; C492280:Slc12a6 protein, mouse; D000096922:K Cl- Cotransporters