A method for bidirectional solution exchange鈥斺€淟iquid bullet鈥?applications of acetylcholine to 伪7 nicotinic receptors

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• 作者：Nikolai Fedorov^a ; ^{Nikolai.Fedorov@Targacept.com} ; Lisa Benson^a ; John D. Graef^a ; Jeremy Hyman^a ; Jill Sollenberger^a ; Fredrik Pettersson^b ; Patrick M. Lippiello^a ; Merouane Bencherif^a
• 关键词：NNR ; neuronal nicotinic receptors ; LB ; liquid bullet ; ACh ; acetylcholine
• 刊名：Journal of Neuroscience Methods
• 出版年：2012
• 期刊代码：33_01650270
• 类别：neu
• 出版时间：30 April, 2012
• 卷：206
• 期：1
• 页码：23-33
• 文件大小：1598 K

摘要

Fast solution exchange techniques have revolutionized the study of synaptic transmission and promise to remain an important neuroscience research tool. Here we provide evidence for the hypothesis that using continuous, rapid transitions through an agonist solution can significantly increase the exchange rate around a cell by reducing the diffusion boundary at the membrane. This novel approach of rapid solution exchange during whole-cell recordings - described as a 鈥渓iquid bullet鈥?(LB) application - takes advantage of a bidirectional solution flow around the cell, allowing for a full solution exchange within a range of several milliseconds. An exchange rate (10-90%rise time) of about 2 ms could be achieved during both agonist application and washout. We recorded whole-cell currents from cells expressing the rapidly desensitizing 伪7 neuronal nicotinic receptor (NNR) subtype that exhibited very fast rise times of around 4-5 ms. We further demonstrated the advantages of a LB application over conventional methods by the ability of this method to elicit concentration-dependent responses for rapidly desensitizing compounds that were not measurable with conventional agonist applications. In addition, we illustrate the utility of this approach for frequency-based assays through fast, repeated agonist applications at frequencies of 1 Hz and 30 Hz. This approach could therefore be useful for the study of rapid agonist-receptor interactions that closely mimic the physiological conditions in the synaptic cleft during bursts of neuronal activity.