Type 1 equilibrative nucleoside transporter regulates astrocyte-specific glial fibrillary acidic protein expression in the striatum. Brain Behav 2014;4(6):903-14
Date
11/05/2014Pubmed ID
25365803Pubmed Central ID
PMC4178301DOI
10.1002/brb3.283Scopus ID
2-s2.0-84911363465 (requires institutional sign-in at Scopus site) 13 CitationsAbstract
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.
Author List
Hinton DJ, Lee MR, Jang JS, Choi DSAuthor
Ryan M. Lee MD Instructor in the Neurology department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AnimalsAstrocytes
Blotting, Western
Cell Line
Corpus Striatum
Equilibrative Nucleoside Transporter 1
Fluorescent Antibody Technique
Gene Expression Profiling
Glial Fibrillary Acidic Protein
Green Fluorescent Proteins
Male
Mice, 129 Strain
Mice, Inbred C57BL
Mice, Knockout
Mice, Transgenic
Microarray Analysis
Nerve Tissue Proteins
Promoter Regions, Genetic
RNA, Messenger
Real-Time Polymerase Chain Reaction
