Characterization of the rat A2A adenosine receptor gene: A 4.8-kb promoter-proximal DNA fragment confers selective expression in the central nervous system

Yi Chao Lee, Chen Li Chien, Chung Nan Sun, Chuen Lin Huang, Nai Kuei Huang, Ming Chang Chiang, Hsing Lin Lai, Yow Sien Lin, Szu Yi Chou, Chi Kuang Leo Wang, Ming Hong Tai, Wen Lin Liao, Teng Nan Lin, Fu Chin Liu, Yijuang Chern

Research output: Contribution to journalArticle

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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 A 2A-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.

Original languageEnglish
Pages (from-to)1786-1796
Number of pages11
JournalEuropean Journal of Neuroscience
Volume18
Issue number7
DOIs
Publication statusPublished - Oct 2003
Externally publishedYes

Fingerprint

Adenosine A2A Receptors
Transgenes
Central Nervous System
DNA
Genes
Corpus Striatum
GABAergic Neurons
Cholinergic Neurons
Enkephalins
5' Flanking Region
Gene Fusion
PC12 Cells
Astrocytes
Adenosine
Neurons
Brain

Keywords

  • Astrocyte
  • Gene expression
  • Neuron
  • Transgenic

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Characterization of the rat A2A adenosine receptor gene : A 4.8-kb promoter-proximal DNA fragment confers selective expression in the central nervous system. / Lee, Yi Chao; Chien, Chen Li; Sun, Chung Nan; Huang, Chuen Lin; Huang, Nai Kuei; Chiang, Ming Chang; Lai, Hsing Lin; Lin, Yow Sien; Chou, Szu Yi; Wang, Chi Kuang Leo; Tai, Ming Hong; Liao, Wen Lin; Lin, Teng Nan; Liu, Fu Chin; Chern, Yijuang.

In: European Journal of Neuroscience, Vol. 18, No. 7, 10.2003, p. 1786-1796.

Research output: Contribution to journalArticle

Lee, YC, Chien, CL, Sun, CN, Huang, CL, Huang, NK, Chiang, MC, Lai, HL, Lin, YS, Chou, SY, Wang, CKL, Tai, MH, Liao, WL, Lin, TN, Liu, FC & Chern, Y 2003, 'Characterization of the rat A2A adenosine receptor gene: A 4.8-kb promoter-proximal DNA fragment confers selective expression in the central nervous system', European Journal of Neuroscience, vol. 18, no. 7, pp. 1786-1796. https://doi.org/10.1046/j.1460-9568.2003.02907.x
Lee, Yi Chao ; Chien, Chen Li ; Sun, Chung Nan ; Huang, Chuen Lin ; Huang, Nai Kuei ; Chiang, Ming Chang ; Lai, Hsing Lin ; Lin, Yow Sien ; Chou, Szu Yi ; Wang, Chi Kuang Leo ; Tai, Ming Hong ; Liao, Wen Lin ; Lin, Teng Nan ; Liu, Fu Chin ; Chern, Yijuang. / Characterization of the rat A2A adenosine receptor gene : A 4.8-kb promoter-proximal DNA fragment confers selective expression in the central nervous system. In: European Journal of Neuroscience. 2003 ; Vol. 18, No. 7. pp. 1786-1796.
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AU - Lee, Yi Chao

AU - Chien, Chen Li

AU - Sun, Chung Nan

AU - Huang, Chuen Lin

AU - Huang, Nai Kuei

AU - Chiang, Ming Chang

AU - Lai, Hsing Lin

AU - Lin, Yow Sien

AU - Chou, Szu Yi

AU - Wang, Chi Kuang Leo

AU - Tai, Ming Hong

AU - Liao, Wen Lin

AU - Lin, Teng Nan

AU - Liu, Fu Chin

AU - Chern, Yijuang

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N2 - 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 A 2A-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.

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