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.
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.
Miyoshi
Yurika
Y
Graduate School of Pharmaceutical Sciences, Kyushu University, 3-1-1 Maidashi, Fukuoka 812-8582, Japan.
Hamase
Kenji
K
Okamura
Tadashi
T
Konno
Ryuichi
R
Kasai
Noriyuki
N
Tojo
Yosuke
Y
Zaitsu
Kiyoshi
K
D000409:Alanine / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000652:urine; D000818:Animals; D001923:Brain Chemistry; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003605:D-Amino-Acid Oxidase / Q000235:genetics / Q000378:metabolism*; D004195:Disease Models, Animal; D007668:Kidney / Q000737:chemistry; D008297:Male; D010179:Pancreas / Q000737:chemistry; D010902:Pituitary Gland; D051381:Rats; D020318:Rats, Long-Evans; D017207:Rats, Sprague-Dawley; D055647:Rats, Transgenic; D017208:Rats, Wistar; D012044:Regression Analysis; D015203:Reproducibility of Results; D012694:Serine / Q000032:analysis* / Q000097:blood / Q000737:chemistry / Q000652:urine; D013237:Stereoisomerism
D012694:Serine; D003605:D-Amino-Acid Oxidase; D000409:Alanine