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首页> 外文期刊>The European Journal of Neuroscience >Second-by-second measurement of acetylcholine release in prefrontal cortex.
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Second-by-second measurement of acetylcholine release in prefrontal cortex.

机译：第二次测量额叶前额叶皮层中乙酰胆碱的释放。

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摘要

Microdialysis has been widely used to measure acetylcholine (ACh) release in vivo and has provided important insights into the regulation of cholinergic transmission. However, microdialysis can be constrained by limited spatial and temporal resolution. The present experiments utilize a microelectrode array (MEA) to rapidly measure ACh release and clearance in anaesthetized rats. The array electrochemically detects, on a second-by-second basis, changes in current selectively produced by the hydrolysis of ACh to choline (Ch) and the subsequent oxidation of choline and hydrogen peroxidase (H(2)O(2)) at the electrode surface. In vitro calibration of the microelectrode revealed linear responses to ACh (R(2) = 0.9998), limit of detection of 0.08 microm, and signal-to-noise ratio of 3.0. The electrode was unresponsive to ascorbic acid (AA), dopamine (DA), or norepinephrine (NE) interferents. In vivo experiments were conducted in prefrontal cortex (PFC) of anaesthetized rats. Pressure ejections of ACh (10 mm; 40 nL) through an adjoining micropipette produced a rapid rise in current, reaching maximum amplitude in approximately 1.0 s and cleared by 80% within 4-11 s. Endogenously released ACh, following local depolarization with KCl (70 mm; 40, 160 nL), was detected at values as low as 0.05 microm. These signals were volume-dependent and cleared within 4-12 s. Finally, nicotine (1.0 mm, 80 nL) stimulated ACh signals. Nicotine-induced signals reflected the hydrolysis of ACh by endogenous acetylcholinesterase (AChE) as inhibition of the enzyme following perfusion with neostigmine (10 microm) attenuated the signal (40-94%). Collectively, these data validate a novel method for rapidly measuring cholinergic transmission in vivo with a spatial and temporal resolution that far exceeds conventional microdialysis.
机译：微透析已被广泛用于体内测量乙酰胆碱（ACh）的释放，并为调节胆碱能传递提供了重要见解。但是，微透析会受到空间和时间分辨率的限制。本实验利用微电极阵列（MEA）快速测量麻醉大鼠中ACh的释放和清除。该阵列以电化学方式每秒检测由ACh水解为胆碱（Ch）以及随后在胆碱和过氧化氢（H（2）O（2））氧化而选择性产生的电流变化。电极表面。微电极的体外校准显示对ACh的线性响应（R（2）= 0.9998），检出限为0.08微米和信噪比为3.0。电极对抗坏血酸（AA），多巴胺（DA）或去甲肾上腺素（NE）干扰物无反应。在麻醉大鼠的前额叶皮层（PFC）中进行了体内实验。通过相邻的微量移液器对ACh（10 mm; 40 nL）进行压力喷射会产生电流的快速上升，在大约1.0 s内达到最大振幅，并在4-11 s内清除80％。在用KCl（70 mm; 40，160 nL）局部去极化后，检测到内源释放的ACh，其值低至0.05微米。这些信号取决于音量，并在4-12 s内清除。最后，尼古丁（1.0 mm，80 nL）刺激了ACh信号。尼古丁诱导的信号反映了内源性乙酰胆碱酯酶（AChE）对ACh的水解，因为在用新斯的明（10 microm）灌注后抑制该酶减弱了信号（40-94％）。总的来说，这些数据验证了一种新颖的方法，该方法可以以远远超过传统微透析的时空分辨率快速测量体内胆碱能传递。

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