Identifying the serotonin transporter signal in Western blot studies of the neurotoxic potential of MDMA and related drugs.
Identifying the serotonin transporter signal in Western blot studies of the neurotoxic potential of MDMA and related drugs.
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在 MDMA 和相关药物潜在神经毒性的蛋白质印迹研究中识别血清素转运蛋白信号。
DOI：
10.1002/syn.20958
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发表时间：
2011
期刊：
影响因子：
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通讯作者：
Ricaurte,George
中科院分区：
文献类型：
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作者：
McLane,MichaelW;McCann,Una;Ricaurte,George
一些已发表的研究质疑了3，4-亚甲基二氧基甲基苯丙胺(MDMA，“摇头丸”)和相关药物(芬氟拉明，对氯苯丙胺)的5-羟色胺神经毒性潜力，这是基于使用定制合成的5-羟色胺转运体(SERT)抗体进行的Western印迹研究的结果，该抗体发现替代苯丙胺治疗后50 kDa蛋白的丰度没有减少。这项研究的目的是使用在这些研究中使用的相同的SERT抗体收集Western印迹数据，但使用阳性和阴性对照来识别SERT蛋白信号。一条63-68 kDa的条带被鉴定为SERT蛋白，该条带具有大鼠脑SERT预期的区域分布，被5，7-DHT减少，在SERT KO动物中不存在。用MDMA和相关药物(Fen、PCA)治疗后，大鼠脑内63-68 kDa条带的丰度显著、持久地下降。因此，当通过使用阳性和阴性对照在Western blotts中鉴定出与SERT蛋白相对应的条带时，替代苯丙胺处理后SERT蛋白丰度的降低很容易被证明。这些数据提供了进一步的证据，证明在接触MDMA和其他替代苯丙胺后，SERT蛋白持续丢失。Synapse，2011年。©2011 Wiley-Liss，Inc.
A number of published studies have questioned the serotonin neurotoxic potential of 3,4‐methylenedioxymethamphetamine (MDMA, “ecstasy”) and related drugs (fenfluramine, p‐chloroamphetamine) based upon results from Western blot studies using a custom synthesized serotonin transporter (SERT) antibody that found no reduction in the abundance of a 50kDa protein after substituted amphetamine treatment. The purpose of this study was to collect Western blot data using the same SERT antibody used in those studies, but with positive and negative controls to identify the SERT protein signal. A 63–68 kDa band that had the regional distribution expected of rat brain SERT, was decreased by 5,7‐DHT, and was absent in SERT KO animals was identified as the SERT protein. Significant, lasting decreases in the abundance of the 63–68 kDa band were evident in the rat brain after treatment with MDMA and related drugs (FEN, PCA). Thus, when the band corresponding to the SERT protein is identified in Western blots through the use of positive and negative controls, reduced abundance of the SERT protein can be readily demonstrated after substituted amphetamine treatment. These data provide further evidence of lasting loss of the SERT protein after exposure to MDMA and other substituted amphetamines. Synapse, 2011. © 2011 Wiley‐Liss, Inc.