title: Characterization of the rat A2A adenosine receptor gene: a 4.8-kb promoter-proximal DNA fragment confers selective expression in the central nervous system.
abstract: We isolated and characterized a 4.8-kb 5' flanking region of the rat A2A adenosine receptor (A2A-R) gene in the present study. Promoter activity was observed with this DNA fragment in PC12 cells and C6 cells which contain endogenous A2A-Rs. A fusion fragment consisting of the 4.8-kb promoter-proximal DNA fragment of the A2A-R gene, and the coding region of lacZ was utilized to produce mice harbouring the fusion gene. In three independent founder lines, proteins and transcripts of the transgene were found in many areas of the central nervous system (CNS), but not in three peripheral tissues examined. Double immunohistochemical analyses revealed that the transgene was coexpressed with endogenous A2A-R and proper neuronal markers in the brain. Specifically, the transgene in the striatum was found in the enkephalin-containing GABAergic neurons and in the cholinergic neurons as was found for the endogenous A2A-R. However, a selectively enriched striatal expression of the transgene was not found as was observed for the endogenous A2A-R. Collectively, the 4.8-kb promoter-proximal DNA fragment of the rat A2A-R gene contains important element(s) to direct its expression in the CNS where functional A2A-R are found, but were not sufficient to confer the highly concentrated expression of the striatal A2A-R. Furthermore, expressions of A2A-R and the transgene were found in both neurons and astrocytes, suggesting that adenosine might mediate its function through A2A-R in both cell types.
authors: Institute of Biomedical Sciences, Academia Sinica, Taipei 11529, Taiwan.
authors: Yi-Chao
authors: YC
authors: Lee
authors: Chen-Li
authors: CL
authors: Chien
authors: Chung-Nan
authors: CN
authors: Sun
authors: Chuen-Lin
authors: CL
authors: Huang
authors: Nai-Kuei
authors: NK
authors: Huang
authors: Ming-Chang
authors: MC
authors: Chiang
authors: Hsing-Lin
authors: HL
authors: Lai
authors: Yow-Sien
authors: YS
authors: Lin
authors: Szu-Yi
authors: SY
authors: Chou
authors: Chi-Kuang Leo
authors: CK
authors: Wang
authors: Ming-Hong
authors: MH
authors: Tai
authors: Wen-Lin
authors: WL
authors: Liao
authors: Teng-Nan
authors: TN
authors: Lin
authors: Fu-Chin
authors: FC
authors: Liu
authors: Yijuang
authors: Y
authors: Chern
mesh_terms: D024506:5' Flanking Region; D000262:Adenylyl Cyclases / Q000494:pharmacology; D000818:Animals; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D015139:Blotting, Southern; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002490:Central Nervous System / Q000033:anatomy & histology / Q000187:drug effects / Q000378:metabolism*; D053938:DNA Fragmentation; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D051379:Mice; D008822:Mice, Transgenic; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D016716:PC12 Cells; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012313:RNA / Q000378:metabolism; D051381:Rats; D043705:Receptor, Adenosine A2A / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014162:Transfection; D014404:Tubulin / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D005904:Glial Fibrillary Acidic Protein; D008869:Microtubule-Associated Proteins; D043705:Receptor, Adenosine A2A; D014404:Tubulin; C462941:beta3 tubulin, mouse; D012313:RNA; D001616:beta-Galactosidase; D000262:Adenylyl Cyclases