Characterization of a Yeast D-Amino Acid Oxidase Microbiosensor for D-Serine Detection in the Central Nervous System

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• 作者：Pierre Pernot ; Jean-Pierre Mothet ; Oleg Schuvailo ; Alexey Soldatkin ; Loredano Pollegioni ; Mirella Pilone ; Marie-Th&eacute ; rè ; se Adeline ; Raymond Cespuglio ; St&eacute ; phane Marinesco
• 刊名：Analytical Chemistry
• 出版年：2008
• 出版时间：March 1, 2008
• 年：2008
• 卷：80
• 期：5
• 页码：1589 - 1597
• 全文大小：386K
• 年卷期：v.80,no.5(March 1, 2008)
• ISSN：1520-6882

文摘

D-Serine is an endogenous ligand for N-methyl-D-aspartate(NMDA) receptors, and alterations in its concentrationhave been related to several brain disorders, especiallyschizophrenia. It is therefore an important target neuromodulator for the pharmaceutical industry. To monitorD-serine levels in vivo, we have developed a microbiosensor based on cylindrical platinum microelectrodes, covered with a membrane of poly-m-phenylenediamine (PPD)and a layer of immobilized D-amino acid oxidase from theyeast Rhodotorula gracilis (RgDAAO). By detecting thehydrogen peroxide produced by enzymatic degradation ofD-serine, this microbiosensor shows a detection limit of16 nM and a mean response time of 2 s. Interferencesby ascorbic acid, uric acid, L-cysteine, and by biogenicamines and their metabolites are rejected at more than97% by the PPD layer. Although several D-amino acidsare potential substrates for RgDAAO, D-serine was theonly endogenous substrate present in sufficient concentration to be detected by our microbiosensor in the centralnervous system. When implanted in the cortex of anesthetized rats, this microbiosensor detected the increasein concentration of D-serine resulting from its diffusionacross the blood-brain barrier after an intraperitonealinjection. This new device will make it possible to investigate in vivo the variations in D-serine concentrationsoccurring under normal and pathological conditions andto assess the pharmacological potency of new drugsdesigned to impact D-serine metabolism.