Papouin Thomas, Haydon Philip G
Department of Neuroscience, Tufts University School of Medicine, Boston, MA, USA.
Bio Protoc. 2018 Jan 20;8(2). doi: 10.21769/BioProtoc.2698.
D-serine is an atypical amino acid present in the mammalian body (most amino acids in the mammalian body are L-isomers) that is mostly known in neuroscience for its role as a co-agonist controlling the N-methyl D-aspartate receptor (NMDAR). D-serine levels are decreased in patients with schizophrenia and this is thought to mediate, at least in part, the hypofunction of NMDARs that is central to the glutamate hypothesis for the etiology of this neuropsychiatric disorder. D-serine detection was first established using high performance liquid chromatography, a costly and complex technique that requires high levels of expertise. But with the increasing interest in this unconventional amino acid, there is an increasing need for easier, cheaper and more accessible detection methods. Here we describe the amperometric, biosensor-based method we employed in a recent publication (Papouin ., 2017b). It allows reliable measurement of D-serine levels from fresh tissue, such as acute brain slices, for concentrations higher than 100 nM, with minimal technical requirements.
D-丝氨酸是一种存在于哺乳动物体内的非典型氨基酸(哺乳动物体内的大多数氨基酸是L-异构体),在神经科学领域,它主要作为一种共同激动剂控制N-甲基-D-天冬氨酸受体(NMDAR)而为人所知。精神分裂症患者体内的D-丝氨酸水平会降低,这被认为至少部分介导了NMDAR功能减退,而NMDAR功能减退是这种神经精神疾病病因学中谷氨酸假说的核心。D-丝氨酸检测最初是使用高效液相色谱法建立的,这是一种成本高昂且复杂的技术,需要高水平的专业知识。但随着对这种非常规氨基酸的兴趣日益增加,对更简便、更便宜且更易获得的检测方法的需求也在不断增加。在此,我们描述了我们在最近一篇出版物(帕普安等人,2017b)中采用的基于生物传感器的安培检测法。它能够可靠地测量来自新鲜组织(如急性脑切片)中浓度高于100 nM的D-丝氨酸水平,且技术要求极低。