title: Type 1 equilibrative nucleoside transporter regulates astrocyte-specific glial fibrillary acidic protein expression in the striatum.
abstract: BACKGROUND
  Adenosine signaling has been implicated in several neurological and psychiatric disorders. Previously, we found that astrocytic excitatory amino acid transporter 2 (EAAT2) and aquaporin 4 (AQP4) are downregulated in the striatum of mice lacking type 1 equilibrative nucleoside transporter (ENT1).
  
  
  METHODS
  To further investigate the gene expression profile in the striatum, we preformed Illumina Mouse Whole Genome BeadChip microarray analysis of the caudate-putamen (CPu) and nucleus accumbens (NAc) in ENT1 null mice. Gene expression was validated by RT-PCR, Western blot, and immunofluorescence. Using transgenic mice expressing enhanced green fluorescence protein (EGFP) under the control of the glial fibrillary acidic protein (GFAP) promoter, we examined EGFP expression in an ENT1 null background.
  
  
  RESULTS
  Glial fibrillary acidic protein was identified as a top candidate gene that was reduced in ENT1 null mice compared to wild-type littermates. Furthermore, EGFP expression was significantly reduced in GFAP-EGFP transgenic mice in an ENT1 null background in both the CPu and NAc. Finally, pharmacological inhibition or siRNA knockdown of ENT1 in cultured astrocytes also reduced GFAP mRNA levels.
  
  
  CONCLUSIONS
  Overall, our findings demonstrate that ENT1 regulates GFAP expression and possibly astrocyte function.
authors.affiliation: Department of Psychiatry and Psychology, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905 ; Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905 ; Neurobiology of Disease Program, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905.
authors.forename: David J
authors.initials: DJ
authors.lastname: Hinton
authors.affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905.
authors.forename: Moonnoh R
authors.initials: MR
authors.lastname: Lee
authors.affiliation: Division of Pulmonary and Critical Care Medicine, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905.
authors.forename: Jin Sung
authors.initials: JS
authors.lastname: Jang
authors.affiliation: Department of Psychiatry and Psychology, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905 ; Department of Molecular Pharmacology and Experimental Therapeutics, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905 ; Neurobiology of Disease Program, Mayo Clinic, College of Medicine Rochester, Minnesota, 55905.
authors.forename: Doo-Sup
authors.initials: DS
authors.lastname: Choi
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000378:metabolism*; D015153:Blotting, Western; D002460:Cell Line; D003342:Corpus Striatum / Q000378:metabolism*; D033721:Equilibrative Nucleoside Transporter 1 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D005904:Glial Fibrillary Acidic Protein; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D008297:Male; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008822:Mice, Transgenic; D046228:Microarray Analysis; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction
chemical_list: D033721:Equilibrative Nucleoside Transporter 1; D005904:Glial Fibrillary Acidic Protein; D009419:Nerve Tissue Proteins; D012333:RNA, Messenger; C498476:SLC29A1 protein, mouse; C485184:enhanced green fluorescent protein; C528900:glial fibrillary astrocytic protein, mouse; D049452:Green Fluorescent Proteins
keywords: Adenosine transporter; ENT1; GFAP; astrocyte; microarray