Ingram S L, Amara S G
Vollum Institute and Howard Hughes Medical Institute, Oregon Health Sciences University, Portland, Oregon 97201, USA.
J Neurosci. 2000 Jan 15;20(2):550-7. doi: 10.1523/JNEUROSCI.20-02-00550.2000.
The dopamine transporter (DAT) exhibits several ionic currents that are either coupled to or uncoupled from the transport of substrate. Second messenger systems have been shown to modulate dopamine (DA) transport, however, the modulation of DAT-associated currents has not been studied in depth. Using the two-electrode voltage-clamp method to record from Xenopus oocytes expressing the human DAT, we examined the effects of arachidonic acid (AA) on membrane currents. AA (10-100 microM) stimulates a novel nonselective cation conductance seen only in oocytes expressing human DA transporter (hDAT). The AA-stimulated conductance is up to 50-fold greater than the current normally elicited by DA, but does not appear to arise from the modulation of previously described hDAT conductances, including the leak current and the current associated with electrogenic transport. In addition, DA dramatically potentiates and cocaine blocks the AA-stimulated DAT current. DA potentiates the AA-induced currents in the absence of sodium and chloride, indicating that these currents arise from processes distinct from those associated with substrate transport. The effects of AA were mimicked by other fatty acids with a rank order of potency correlated with their degree of unsaturation, suggesting that AA directly stimulates the novel cation current. Therefore, AA stimulation of this DAT-associated conductance may provide a novel mechanism for modulation of neuronal signaling.
多巴胺转运体(DAT)表现出几种离子电流,这些电流要么与底物转运偶联,要么与之解偶联。第二信使系统已被证明可调节多巴胺(DA)转运,然而,与DAT相关电流的调节尚未得到深入研究。我们使用双电极电压钳法记录表达人DAT的非洲爪蟾卵母细胞的电流,研究了花生四烯酸(AA)对膜电流的影响。AA(10 - 100微摩尔)刺激了一种仅在表达人多巴胺转运体(hDAT)的卵母细胞中可见的新型非选择性阳离子电导。AA刺激的电导比DA正常引发的电流大50倍,但似乎并非源于对先前描述的hDAT电导的调节,包括漏电流和与电转运相关的电流。此外,DA显著增强而可卡因阻断AA刺激的DAT电流。在没有钠和氯的情况下,DA增强AA诱导的电流,表明这些电流源于与底物转运不同的过程。其他脂肪酸模拟了AA的作用,其效力顺序与其不饱和度相关,表明AA直接刺激了新型阳离子电流。因此,AA对这种与DAT相关的电导的刺激可能为神经元信号调节提供一种新机制。