Journal
NEUROSCIENCE LETTERS
Volume 324, Issue 2, Pages 101-104Publisher
ELSEVIER SCI IRELAND LTD
DOI: 10.1016/S0304-3940(02)00184-2
Keywords
D-amino acid oxidase; astrocyte; D-serine; N-methyl-D-aspartate receptor; reverse transcriptase-polymerase chain reaction; cell separation; CG-4; C6
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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. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.
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