Geiger Brenda M, Frank Lauren E, Caldera-Siu Angela D, Pothos Emmanuel N
Department of Pharmacology and Experimental Therapeutics, Tufts University School of Medicine, USA.
J Vis Exp. 2008 Oct 6(20):880. doi: 10.3791/880.
The ability to measure extracellular basal levels of neurotransmitters in the brain of awake animals allows for the determination of effects of different systemic challenges (pharmacological or physiological) to the CNS. For example, one can directly measure how the animal's midbrain dopamine projections respond to dopamine-releasing drugs like d-amphetamine or natural stimuli like food. In this video, we show you how to implant guide cannulas targeting specific sites in the rat brain, how to insert and implant a microdialysis probe and how to use high performance liquid chromatography coupled with electrochemical detection (HPLC-EC) to measure extracellular levels of oxidizable neurotransmitters and metabolites. Local precise introduction of drugs through the microdialysis probe allows for refined work on site specificity in a compound s mechanism of action. This technique has excellent anatomical and chemical resolution but only modest time resolution as microdialysis samples are usually processed every 20-30 minutes to ensure detectable neurotransmitter levels. Complementary ex vivo tools (i.e., slice and cell culture electrophysiology) can assist with monitoring real-time neurotransmission.
测量清醒动物大脑中神经递质细胞外基础水平的能力,有助于确定不同的全身性刺激(药理学或生理学刺激)对中枢神经系统的影响。例如,人们可以直接测量动物中脑多巴胺投射对诸如右旋苯丙胺等多巴胺释放药物或食物等自然刺激的反应。在本视频中,我们向您展示如何在大鼠大脑中靶向特定部位植入引导套管,如何插入和植入微透析探针,以及如何使用高效液相色谱与电化学检测联用技术(HPLC-EC)来测量可氧化神经递质和代谢物的细胞外水平。通过微透析探针在局部精确引入药物,有助于在化合物作用机制方面进行更精细的位点特异性研究。这项技术具有出色的解剖学和化学分辨率,但时间分辨率一般,因为微透析样本通常每20 - 30分钟处理一次,以确保可检测到神经递质水平。补充性的离体工具(即切片和细胞培养电生理学)可协助监测实时神经传递。