Effects of ethanol on extracellular levels of adenosine in the basal forebrain: an in vivo microdialysis study in freely behaving rats.
Effects of ethanol on extracellular levels of adenosine in the basal forebrain: an in vivo microdialysis study in freely behaving rats.
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DOI：
10.1111/j.1530-0277.2010.01153.x
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发表时间：
2010-05
期刊：
影响因子：
--
通讯作者：
Thakkar MM
中科院分区：
文献类型：
--
作者：
Sharma R;Engemann SC;Sahota P;Thakkar MM
Adenosine is implicated to play a pivotal role in mediating many neuronal responses to ethanol. While in vitro studies performed in cell culture have demonstrated that acute ethanol exposure increases extracellular adenosine levels, this effect has not been demonstrated, in vivo, in the brain. We performed an in vivo microdialysis study to examine the effects of local ethanol perfusion on extracellular levels of adenosine in the basal forebrain. Under sterile conditions and using a standard surgical protocol, adult male Sprague-Dawley rats were implanted with unilateral microdialysis guide cannula targeted towards the basal forebrain. Following post-operative recovery, the microdialysis probe was inserted. After allowing at least 12–16 hr for probe insertion recovery, the experiment was begun. Artificial cerebrospinal fluid was perfused (0.7 μL/min) for 80 min and 4 × 20 min pre-ethanol baseline samples were collected. Subsequently, 30, 100 and 300mM doses of ethanol were perfused. Each ethanol dose was perfused for 80 min and 4 × 20 min samples were collected. Finally, aCSF was perfused and 4 × 20 post-ethanol samples were collected. Adenosine in the microdialysate was separated and measured with HPLC coupled with an UV detector. On completion, the animals were euthanized, brain removed and processed for histology. Local ethanol perfusion in the basal forebrain produced a significant increase in extracellular adenosine with the highest dose of 300 mM ethanol producing a 4 fold increase. Cresyl violet (Nissl) staining did not indicate any toxic damage in the area surrounding the probe tip. Choline acetyltransferase immunohistochemistry revealed that all microdialysis probe sites were localized in the basal forebrain. Our study is the first to demonstrate that ethanol acts directly in the brain to increase extracellular adenosine.
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