title: Genetic and functional studies of a missense variant in a glutamate transporter, SLC1A3, in Tourette syndrome.
abstract: OBJECTIVE
  Abnormalities in neurotransmission within the cortico-striatal-thalamo-cortical circuitry are implicated in the pathogenesis of Tourette syndrome. Glutamate is a major excitatory neurotransmitter and an important member in the cortico-striatal-thalamo-cortical circuitry. To explore the role of glutamatergic neurotransmission in genetic susceptibility of Tourette syndrome, we carried out the genetic and functional characterization of sequence variants in SLC1A3 gene, which encodes the main glutamate transporter in astrocytes in individuals with well-characterized Tourette syndrome (n=256) and normal controls (n=224).
  
  
  METHODS
  Exon-containing regions of SLC1A3 gene were screened using capillary electrophoresis-single strand conformation polymorphism followed by direct sequencing. Sequence variants were genotyped by restriction enzyme digestion and studied using glutamate uptake assay and membrane protein pull-down for transporter function.
  
  
  RESULTS
  A missense variant involving a highly conserved residue, E219D, was identified in 11 heterozygous individuals with Tourette syndrome and four in the controls. The allele frequency for E219D was 2.4 folds higher in the Tourette syndrome (0.022) compared with the control cohort (0.009) although the difference did not reach statistical significance in the current cohorts (P=0.09). A H-glutamate-uptake assay showed that E219D conveys a significant increase (1.66 fold) in the SLC1A3-mediated glutamate uptake in HEK293 cells. A biotin-mediated membrane pull-down analysis showed a similar increase (1.5 fold) of mutant SLC1A3 protein in the membrane fraction of transfected HEK293 cells compared with that in the wild type controls.
  
  
  CONCLUSIONS
  These results indicate that E219D is a functional SLC1A3 variant that is presented in a small number of individuals with Tourette syndrome. Further studies on possible changes in glutamate transport in the pathogenesis of Tourette syndrome are warranted.
authors.affiliation: McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
authors.forename: Abby
authors.initials: A
authors.lastname: Adamczyk
authors.forename: Colin D
authors.initials: CD
authors.lastname: Gause
authors.forename: Rita
authors.initials: R
authors.lastname: Sattler
authors.forename: Svetlana
authors.initials: S
authors.lastname: Vidensky
authors.forename: Jeffery D
authors.initials: JD
authors.lastname: Rothstein
authors.forename: Harvey
authors.initials: H
authors.lastname: Singer
authors.forename: Tao
authors.initials: T
authors.lastname: Wang
mesh_terms: D000483:Alleles; D001483:Base Sequence; D001519:Behavior; D002648:Child; D002675:Child, Preschool; D017124:Conserved Sequence / Q000235:genetics; D004252:DNA Mutational Analysis; D019075:Electrophoresis, Capillary; D027341:Excitatory Amino Acid Transporter 1 / Q000235:genetics*; D005787:Gene Frequency / Q000235:genetics; D020022:Genetic Predisposition to Disease; D018698:Glutamic Acid / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D007223:Infant; D008969:Molecular Sequence Data; D020125:Mutation, Missense / Q000235:genetics*; D018807:Polymorphism, Single-Stranded Conformational; D013347:Subcellular Fractions / Q000378:metabolism; D005879:Tourette Syndrome / Q000235:genetics*
chemical_list: D027341:Excitatory Amino Acid Transporter 1; C493394:SLC1A3 protein, human; D018698:Glutamic Acid