Department of Neurophysiology and Brain Sciences, Nagoya City University Graduate School of Medical Sciences, Nagoya, Japan.
Intermedical Co., Ltd, Nagoya, Japan.
J Neurosci Res. 2018 May;96(5):817-827. doi: 10.1002/jnr.24193. Epub 2017 Nov 1.
Fast-scan cyclic voltammetry (FSCV) is an established method for measuring dopamine (DA) levels in the brain in real time. However, it is difficult to discriminate DA from other monoamines such as serotonin (5-hydroxytryptamine, 5-HT) and norepinephrine (NE). We report a novel DA-specific biosensor consisting of a carbon-fiber electrode coated with an ion-exchange membrane, a layer containing monoamine oxidase B, and a cellulose membrane. We performed FSCV using the probe to monitor the amount of DA in vitro and in vivo. First, we measured currents in vitro in phosphate-buffered saline as we added one micromole each of DA, 5-HT, and NE. The results confirmed that the biosensor selectively detected DA. Next, we implanted the probe in the striatum of male rats to investigate whether it could selectively detect changes in the DA content in vivo. The probe detected both the tonic change induced by methamphetamine administration and the phasic change induced by electrical stimulation of the medial forebrain bundle. In contrast, the electrode in the 6-hydroxydopamine-lesioned striatum did not respond to systemic selective serotonin or serotonin/norepinephrine reuptake inhibitors, confirming its selectivity. Furthermore, the probe in the striatum could still detect changes in the DA level 1 week after electrode implantation. The results suggest that the novel biosensor can measure real-time changes in DA levels in vivo with a relatively high signal-to-noise ratio.
快速扫描循环伏安法(FSCV)是一种实时测量大脑中多巴胺(DA)水平的成熟方法。然而,它很难将 DA 与其他单胺类物质(如 5-羟色胺(5-HT)和去甲肾上腺素(NE))区分开来。我们报告了一种新型的 DA 特异性生物传感器,它由涂有离子交换膜的碳纤维电极、含有单胺氧化酶 B 的层和纤维素膜组成。我们使用该探针进行 FSCV 以监测体外和体内的 DA 量。首先,我们在磷酸盐缓冲盐溶液中进行了体外电流测量,同时加入了 1 微摩尔的 DA、5-HT 和 NE。结果证实了该生物传感器对 DA 具有选择性检测能力。接下来,我们将探针植入雄性大鼠的纹状体中,以研究其是否可以选择性地检测体内 DA 含量的变化。该探针可以检测到由苯丙胺给药引起的紧张性变化和由中脑边缘束电刺激引起的阶段性变化。相比之下,在 6-羟多巴胺损伤的纹状体中的电极对全身选择性 5-HT 或 5-HT/去甲肾上腺素再摄取抑制剂没有反应,证实了其选择性。此外,植入电极 1 周后,纹状体中的探针仍能检测到 DA 水平的变化。这些结果表明,新型生物传感器可以以相对较高的信噪比测量体内 DA 水平的实时变化。