Simultaneous two-dimensional HPLC determination of free D-serine and D-alanine in the brain and periphery of mutant rats lacking D-amino-acid oxidase
Simultaneous two-dimensional HPLC determination of free D-serine and D-alanine in the brain and periphery of mutant rats lacking D-amino-acid oxidase
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DOI：
10.1016/j.jchromb.2010.08.024
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发表时间：
2011-11-01
影响因子：
3
通讯作者：
Zaitsu, Kiyoshi
中科院分区：
文献类型：
--
作者：
Miyoshi, Yurika;Hamase, Kenji;Zaitsu, Kiyoshi
设计并验证了微孔ods柱与窄孔对端选择性柱相结合的全自动二维高效液相色谱系统，测定了缺乏d -氨基酸氧化酶（DAO）的Long-Evans agouti/SENDAI （LEA/Sen）大鼠各组织和生理液中d -丝氨酸（D-Ser）和d -丙氨酸（D-Ala）的含量。日内、日间精密度小于4.3%，准确度在99.9% ~ 104%之间。LEA/Sen大鼠的肾脏缺乏DAO，预计是一种新的缺乏DAO的突变动物，然而，LEA/Sen大鼠中d -氨基酸的含量尚未被研究。本研究测定了LEA/Sen大鼠7个脑组织、4个外周组织、血浆和尿液中n -甲基- d -天冬氨酸（NMDA）受体的神经调节剂D-Ser和D-Ala的内在含量，并与DAO活性正常的对照组（Wistar和SD）进行了比较。除额叶区外，LEA/Sen大鼠各组织及生理体液中D-Ser水平均显著高于Wistar和SD大鼠。与Wistar和SD大鼠相比，LEA/Sen大鼠组织和生理体液中D-Ala的含量显著增加。这些结果表明，大鼠组织中D-Ser和D-Ala的内在量受DAO的调节，LEA/Sen大鼠可用于研究与DAO相关的NMDA受体相关疾病。(C) 2010 Elsevier B.V.版权所有
A fully automated two-dimensional HPLC system combining a microbore-ODS column and a narrowbore-enantioselective column was designed and validated, and the amounts of D-serine (D-Ser) and D-alanine (D-Ala) in various tissues and physiological fluids of Long-Evans agouti/SENDAI (LEA/Sen) rats lacking D-amino-acid oxidase (DAO) were determined. Intra- and inter-day precision was less than 4.3% and accuracy ranged between 99.9 and 104%. LEA/Sen rats were reported to lack DAO in their kidneys and expected to be a novel mutant animal lacking DAO, however, the amounts of D-amino acids in the LEA/Sen rats have not been investigated. In the present study, the intrinsic amounts of D-Ser and D-Ala, which are neuromodulators of the N-methyl-D-aspartate (NMDA) receptors, were determined in seven brain tissues, four peripheral tissues, plasma and urine of the LEA/Sen rats and compared to those of the control (Wistar and SD) rats having normal DAO activity. The levels of D-Ser in the tissues and physiological fluids of the LEA/Sen rats were significantly higher than those of the Wistar and SD rats except for the frontal brain regions. Concerning D-Ala, the amounts in the tissues and physiological fluids of the LEA/Sen rats were drastically increased compared to those of the Wistar and SD rats. These results indicate that the intrinsic amounts of D-Ser and D-Ala in the tissues of rats are regulated by DAO, and that LEA/Sen rats would be useful for the study of NMDA receptor-related diseases in which DAO is implicated. (C) 2010 Elsevier B.V. All rights reserved.