Isoflurane induces dopamine transporter trafficking into the cell cytoplasm
Isoflurane induces dopamine transporter trafficking into the cell cytoplasm
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DOI：
10.1002/syn.20037
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发表时间：
2004-08-01
期刊：
影响因子：
2.3
通讯作者：
Denson, DD
中科院分区：
文献类型：
--
作者：
Byas-Smith, MG;Li, J;Denson, DD
先前的研究表明，在深度异氟烷麻醉期间，猴纹状体中多巴胺转运蛋白（DAT）（2 β-甲氧羰基-3 β-（4-氯苯基）-8-（2-（18）F-氟乙基）去甲托烷）的选择性亲水性正电子发射断层扫描放射性示踪剂的结合减少。免疫组织化学实验表明，但没有证明，异氟烷诱导细胞表面DAT的减少。目前的调查进行了定量证明异氟烷诱导的DAT内化使用快速和相对简单的生化技术在人胚肾（HEK-293）细胞稳定表达人DAT（h-DAT）蛋白。生物素化后进行蛋白质印迹分析，以确定在对照条件下和存在临床相关浓度异氟烷的情况下DAT细胞表面表达的变化程度。异氟醚处理30分钟导致细胞表面h-DAT的量非常显著地减少（对照组的21 +/-15%; P
Previous investigations have shown that the binding of a selective hydrophilic positron emission tomography radiotracer for the dopamine transporter (DAT) (2beta-carbomethoxy-3beta-(4-chlorophenyl)-8-(2-(18)F-fluoroethyl)nortropane) decreased in monkey striatum during deep isoflurane anesthesia. Immunohistochemistry experiments suggested but did not prove that isoflurane induced a decrease in cell surface DAT. The present investigation was undertaken to demonstrate quantitatively the isoflurane-induced internalization of DAT using a rapid and relatively uncomplicated biochemical technique in human embryonic kidney (HEK-293) cells stably expressing the human DAT (h-DAT) protein. Biotinylation followed by Western blot analysis was used to determine the extent of change in cell surface expression of the DAT under control conditions and in the presence of a clinically relevant concentration of isoflurane. Isoflurane treatment for 30 min resulted in a highly significant decrease in the amount of h-DAT on the cell surface (21 +/- 15% of control; P