Real-time monitoring of intracellular cAMP during acute ethanol exposure.
Real-time monitoring of intracellular cAMP during acute ethanol exposure.
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DOI：
10.1111/acer.12133
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发表时间：
2013-09
期刊：
影响因子：
--
通讯作者：
Yoshimura M
中科院分区：
文献类型：
--
作者：
Gupta R;Qualls-Creekmore E;Yoshimura M
In previous studies we have shown that ethanol enhances the activity of Gs-stimulated membrane-bound adenylyl cyclase (AC). The effect is AC isoform specific and the type 7 AC (AC7) is most responsive to ethanol. In this study, we employed a fluorescence resonance energy transfer (FRET) based cAMP sensor, Epac1-camps, to examine real-time temporal dynamics of ethanol effects on cAMP concentrations. To our knowledge, this is the first report on real-time detection of the ethanol effect on intracellular cAMP. Hela cells were transfected with Epac1-camps, dopamine D1A receptor, and one isoform of AC (AC7 or AC3). Fluorescent images were captured using a specific filter set for cyan fluorescent protein (CFP), yellow fluorescent protein (YFP), and FRET, respectively and FRET intensity was calculated on a pixel-by-pixel basis to examine changes in cAMP. During 2-minute stimulation with dopamine (DA), the cytoplasmic cAMP level quickly increased, then decreased to a plateau, where the cAMP level was higher than the level prior to stimulation with DA. Ethanol concentration dependently increased cytoplasmic cAMP in cells transfected with AC7, while ethanol did not have effect on cells transfected with AC3. Similar trends were observed for cAMP at the plasma membrane and in the nucleus during 2-minute stimulation with DA. Unexpectedly, when cells expressing AC7 were stimulated with DA or other Gs protein-coupled receptor’s ligand plus ethanol for 5 seconds, ethanol reduced cAMP concentration. These results suggest that ethanol has two opposing effects on the cAMP generating system in an AC isoform specific manner, the enhancing effect on AC activity and the short lived inhibitory effect. Thus, ethanol may have a different effect on cAMP depending on not only AC isoform but also the duration of exposure.
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DOI：
10.1111/j.1530-0277.1998.tb05903.x
发表时间：
1998-12-01
期刊：
ALCOHOL-CLINICAL AND EXPERIMENTAL RESEARCH
影响因子：
--
作者：
Menninger, JA;Barón, AE;Tabakoff, B
通讯作者：
Tabakoff, B
DOI：
10.1111/j.1440-1681.2008.05020.x
发表时间：
2008-11-01
影响因子：
2.9
作者：
Iancu, Radu V.;Ramamurthy, Gopalakrishnan;Harvey, Robert D.
通讯作者：
Harvey, Robert D.
DOI：
10.1111/j.1530-0277.1999.tb04160.x
发表时间：
1999-04-01
影响因子：
3.2
作者：
Ratsma, JE;Gunning, WB;Schoffelmeer, ANM
通讯作者：
Schoffelmeer, ANM
影响因子：
64.8
作者：
DEARRY, A;GINGRICH, JA;CARON, MG
通讯作者：
CARON, MG
影响因子：
8.4
作者：
Nikolaev, VO;Lohse, MJ
通讯作者：
Lohse, MJ