Validation of a Reversed Phase UPLC-MS/MS Method to Determine Dopamine Metabolites and Oxidation Intermediates in Neuronal Differentiated SH-SY5Y Cells and Brain Tissue.,ACS Chemical Neuroscience

当前位置： X-MOL 学术 › ACS Chem. Neurosci. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Validation of a Reversed Phase UPLC-MS/MS Method to Determine Dopamine Metabolites and Oxidation Intermediates in Neuronal Differentiated SH-SY5Y Cells and Brain Tissue.
ACS Chemical Neuroscience ( IF 4.1 ) Pub Date : 2020-08-06 , DOI: 10.1021/acschemneuro.0c00336
Marta Gonzalez-Sepulveda ₁ , Ariadna Laguna ₁ , Iria Carballo-Carbajal ₁ , Jordi Galiano-Landeira ₁ , Jordi Romero-Gimenez ₁ , Thais Cuadros ₁ , Annabelle Parent ₁ , Nuria Peñuelas ₁ , Joan Compte ₁ , Alba Nicolau ₁ , Camille Guillard-Sirieix ₁ , Helena Xicoy ₁ , Jumpei Kobayashi ₁ , Miquel Vila _{1,

2,

3}

Affiliation

Dopamine is a key neurotransmitter in the pathophysiology of various neurological disorders such as addiction or Parkinson’s disease. Disturbances in its metabolism could lead to dopamine accumulation in the cytoplasm and an increased production of o-quinones and their derivatives, which have neurotoxic potential and act as precursors in neuromelanin synthesis. Thus, quantification of the dopaminergic metabolism is essential for monitoring changes that may contribute to disease development. Here, we developed and validated an UPLC-MS/MS method to detect and quantify a panel of eight dopaminergic metabolites, including the oxidation product aminochrome. Our method was validated in differentiated SH-SY5Y cells and mouse brain tissue and was then employed in brain samples from humans and rats to ensure method reliability in different matrices. Finally, to prove the biological relevance of our method, we determined metabolic changes in an in vitro cellular model of dopamine oxidation/neuromelanin production and in human postmortem samples from Parkinson’s disease patients. The current study provides a validated method to simultaneously monitor possible alterations in dopamine degradation and o-quinone production pathways that can be applied to in vitro and in vivo experimental models of neurological disorders and human brain samples.

中文翻译：

反相UPLC-MS / MS方法确定神经元分化的SH-SY5Y细胞和脑组织中多巴胺代谢产物和氧化中间体的验证。

多巴胺是各种神经系统疾病（例如成瘾或帕金森氏病）的病理生理学中的关键神经递质。代谢紊乱可能导致多巴胺在细胞质中积累并增加o的产生-醌及其衍生物，具有神经毒性潜力，可作为神经黑色素合成的前体。因此，定量多巴胺能代谢对于监测可能有助于疾病发展的变化至关重要。在这里，我们开发并验证了一种UPLC-MS / MS方法，用于检测和定量检测包括氧化产物氨基色素在内的八种多巴胺能代谢产物。我们的方法在分化的SH-SY5Y细胞和小鼠脑组织中得到验证，然后在人和大鼠的脑样本中使用，以确保方法在不同基质中的可靠性。最后，为了证明我们方法的生物学相关性，我们在体外确定了代谢变化多巴胺氧化/ neuromelanin产生的细胞模型以及帕金森氏病患者的人体死后样品中的模型。当前的研究提供了一种经过验证的方法，可以同时监测多巴胺降解和邻醌生成途径的可能变化，这些变化可以应用于神经系统疾病和人脑样本的体外和体内实验模型。

更新日期：2020-09-02

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11