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通过 DNA 靶向扩增检测帕金森病模型中的多巴胺的表面增强拉曼光谱检测。

Surface-enhanced Raman spectroscopy detection of dopamine by DNA Targeting amplification assay in Parkisons's model.

机构信息

Division of Food Bioscience, Konkuk University, Chungju-si 380-701, South Korea.

Department of Biotechnology, Konkuk University, Chungju-si 380-701, South Korea.

出版信息

Biosens Bioelectron. 2015 May 15;67:739-46. doi: 10.1016/j.bios.2014.10.049. Epub 2014 Oct 23.

Abstract

Dopamine is a potent neuromodulator in the brain that influences a variety of motivated behaviors and is involved in several neurologic diseases. We evaluated a bio-barcode amplification assay for its ability to detect dopamine in a mouse model with and without prior administration of the neurotoxin 1-methyl-4-phenyl-1, 2, 3, 6-tetrahydropyridine (MPTP). Our approach uses a combination of DNA barcodes and bead-based immunoassays for detecting neurotransmitters with surface-enhanced Raman spectroscopy (SERS). This method relies on a gold nanoplate with adsorbed antibodies and gold nanoparticles that are encoded with DNA and antibodies that can sandwich the target protein captured by the nanoparticle-bound antibodies. C57BL/6 mice were infused intranasally with MPTP (25mg/kg/day) over 7 consecutive days. At 7 and 21 days after the last administration of MPTP, dopamine was found by western blot analysis to have decreased in the midbrain by 37.44% and 92.95%, respectively. Furthermore, the Raman intensity of dopamine in the midbrains of MPTP-treated mice decreased by 56.77% and 61.12% on days 7 and 21, respectively. Our results demonstrate that the concentration of dopamine in midbrain and striatum of MPTP-treated mice can be easily detected using the bio-barcode assay, which is a rapid, high-throughput screening tool for detecting neurotransmitters.

摘要

多巴胺是大脑中一种有效的神经调节剂,它影响多种动机行为,并参与多种神经疾病。我们评估了一种生物条形码放大测定法,以检测经神经毒素 1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)预处理和未预处理的小鼠模型中的多巴胺。我们的方法结合了 DNA 条码和基于珠的免疫测定法,用于使用表面增强拉曼光谱(SERS)检测神经递质。该方法依赖于吸附有抗体的金纳米板和编码有 DNA 的金纳米粒子,以及可以夹住由纳米粒子结合的抗体捕获的靶蛋白的抗体。C57BL/6 小鼠经鼻内注射 MPTP(25mg/kg/天),连续 7 天。在最后一次给予 MPTP 后的 7 天和 21 天,通过 Western blot 分析发现中脑多巴胺分别减少了 37.44%和 92.95%。此外,MPTP 处理小鼠的中脑多巴胺的拉曼强度在第 7 天和第 21 天分别减少了 56.77%和 61.12%。我们的结果表明,使用生物条形码测定法可以轻松检测 MPTP 处理小鼠中脑和纹状体多巴胺的浓度,这是一种用于检测神经递质的快速、高通量筛选工具。

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