Department of Pharmacotherapy and Translational Research, University of Florida, Gainesville, Florida 32610-0486, USA.
J Neurochem. 2010 Jul;114(1):142-9. doi: 10.1111/j.1471-4159.2010.06738.x. Epub 2010 Apr 6.
The organic cation transporter (OCT) 3 is widely expressed in various organs in humans, and involved in the disposition of many exogenous and endogenous compounds. Several lines of evidence have suggested that OCT3 expressed in the brain plays an important role in the regulation of neurotransmission. Relative to wild-type (WT) animals, Oct3 knockout (KO) mice have displayed altered behavioral and neurochemical responses to psychostimulants such as amphetamine (AMPH) and methamphetamine. In the present study, both in vitro and in vivo approaches were utilized to explore potential mechanisms underlying the disparate neuropharmacological effects observed following AMPH exposure in Oct3 KO mice. In vitro uptake studies conducted in OCT3 transfected cells indicated that dextroamphetamine (d-AMPH) is not a substrate of OCT3. However, OCT3 was determined to be a high-capacity and low-affinity transporter for the neurotransmitters dopamine (DA), norepinephrine (NE), and serotonin (5-HT). Inhibition studies demonstrated that d-AMPH exerts relatively weak inhibitory effects on the OCT3-mediated uptake of DA, NE, 5-HT, and the model OCT3 substrate 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide. The IC(50) values were determined to be 41.5 +/- 7.5 and 24.1 +/- 7.0 microM for inhibiting DA and 5-HT uptake, respectively, while 50% inhibition of NE and 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide uptake was not achieved by even the highest concentration of d-AMPH applied (100 microM). Furthermore, the disposition of d-AMPH in various tissues including the brain, liver, heart, kidney, muscle, intestine, spleen, testis, uterus, and plasma were determined in both male and female Oct3 KO and WT mice. No significant difference was observed between either genotypes or sex in all tested organs and tissues. Our findings suggest that OCT3 is not a prominent factor influencing the disposition of d-AMPH. Additionally, based upon the inhibitory potency observed in vitro, d-AMPH is unlikely to inhibit the uptake of monoamines mediated by OCT3 in the brain. Differentiated neuropharmacological effects of AMPHs noted between Oct3 KO and WT mice appear to be due to the absence of Oct3 mediated uptake of neurotransmitters in the KO mice.
有机阳离子转运体(OCT)3 在人体的各种器官中广泛表达,参与许多外源性和内源性化合物的处置。有几条证据表明,大脑中表达的 OCT3 在调节神经递质方面发挥着重要作用。与野生型(WT)动物相比,Oct3 敲除(KO)小鼠对安非他命(AMPH)和甲基苯丙胺等精神兴奋剂的行为和神经化学反应发生改变。在本研究中,利用体外和体内方法探讨了在 Oct3 KO 小鼠暴露于 AMPH 后观察到的不同神经药理学效应的潜在机制。在 OCT3 转染细胞中进行的体外摄取研究表明,右旋苯丙胺(d-AMPH)不是 OCT3 的底物。然而,OCT3 被确定为多巴胺(DA)、去甲肾上腺素(NE)和 5-羟色胺(5-HT)等神经递质的高容量、低亲和力转运体。抑制研究表明,d-AMPH 对 OCT3 介导的 DA、NE、5-HT 和模型 OCT3 底物 4-(4-(二甲氨基)苯乙烯基)-N-甲基吡啶鎓碘化物摄取的抑制作用相对较弱。IC50 值分别为 41.5 +/- 7.5 和 24.1 +/- 7.0 microM,分别抑制 DA 和 5-HT 摄取,而即使应用最高浓度的 d-AMPH(100 microM)也不能达到 50%抑制 NE 和 4-(4-(二甲氨基)苯乙烯基)-N-甲基吡啶鎓碘化物摄取。此外,在雄性和雌性 Oct3 KO 和 WT 小鼠中,确定了 d-AMPH 在包括大脑、肝脏、心脏、肾脏、肌肉、肠道、脾脏、睾丸、子宫和血浆在内的各种组织中的分布。在所有测试的器官和组织中,两种基因型或性别之间均无显著差异。我们的研究结果表明,OCT3 不是影响 d-AMPH 处置的重要因素。此外,根据体外观察到的抑制效力,d-AMPH 不太可能抑制大脑中由 OCT3 介导的单胺摄取。在 Oct3 KO 和 WT 小鼠之间观察到的 AMPH 神经药理学作用的差异似乎是由于 KO 小鼠中缺乏 Oct3 介导的神经递质摄取所致。