Gene expression of D-amino acid oxidase in cultured rat astrocytes: regional and cell type specific expression.
Neuromodulative free D-serine is present in mammalian brain, and localized to type-2 astrocytes in culture. D-amino acid oxidase (DAO) is a flavoenzyme that catalyzes D-amino acids. We examined the DAO gene expression in cultured rat astrocytes by reverse transcriptase-polymerase chain reaction. We established a method to prepare highly purified culture of type-1 and type-2 astrocytes from any brain region. This method utilizes combination of cell type specific separation by shaking and subsequent purification by immunopanning or treatment with cytosine arabinoside. We detected higher DAO gene expression in type-1 astrocyte cultures from cerebellum than that from cerebral cortex. In cerebellum, we observed higher DAO expression in type-1 astrocyte cultures than that in type-2. We also revealed that DAO expression in C6, corresponding to type-1 astrocyte, was higher than that in CG-4 derived type-2 astrocytes.
Urai
Yoshiteru
Y
Department of Gene Regulatorics, Institute for Enzyme Research, The University of Tokushima, Tokushima 770-8503, Japan.
Jinnouchi
Osamu
O
Kwak
Kyung Tak
KT
Suzue
Atsuhiko
A
Nagahiro
Shinji
S
Fukui
Kiyoshi
K
D000818:Animals; D000954:Antigens, Surface / Q000378:metabolism; D001253:Astrocytes / Q000166:cytology* / Q000201:enzymology*; D001921:Brain / Q000166:cytology* / Q000201:enzymology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D002531:Cerebellum / Q000166:cytology / Q000201:enzymology; D002540:Cerebral Cortex / Q000166:cytology / Q000201:enzymology; D003561:Cytarabine; D003605:D-Amino-Acid Oxidase / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D005904:Glial Fibrillary Acidic Protein; D012333:RNA, Messenger; C046045:Ran 2 protein, rat; D003561:Cytarabine; D003605:D-Amino-Acid Oxidase