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星形胶质细胞培养物对[U-(13)C]天冬氨酸的代谢:培养基的核磁共振分析

Metabolism of [U-(13)C]aspartate by astroglial cultures: nuclear magnetic resonance analysis of the culture media.

作者信息

Murín Radovan, Mohammadi Ghasem, Kowtharapu Bhavani S, Leibfritz Dieter, Hamprecht Bernd

机构信息

Interfaculty Institute for Biochemistry, University of Tuebingen, Hoppe-Seyler-Str 4, 72076 Tuebingen, Germany.

出版信息

Neurochem Res. 2010 Dec;35(12):2053-61. doi: 10.1007/s11064-010-0326-9. Epub 2010 Nov 25.

Abstract

In brain the amino acid L-aspartate serves roles as: (1) putative transmitter, (2) protein precursor, (3) donor of atoms for the biosynthesis of pyrimidine and purine bases, and (4) fuel for energy metabolism. Astrocytes dominate aspartate clearance in brain, and in culture they take up aspartate and quickly metabolize it. In brain, only astrocytes were shown to express the enzymes for de novo pyrimidine biosynthesis. To gain more details about the spectrum of metabolites generated from aspartate and subsequently released by cultured astrocytes a (13)C-nuclear magnetic resonance analysis was performed of [U-(13)C]aspartate supplemented incubation media exposed to astroglial cultures. The results show that astrocytes readily metabolize aspartate and release into their culture media (13)C-isotopomers of lactate, glutamine, citrate and alanine. Despite the presence in astroglial cells of two tandem enzymes of pyrimidine biosynthesis and their mRNAs, pyrimidine nucleotide-related heterocyclic compounds such as dihydroorotate and orotate could not be detected in the culture media.

摘要

在大脑中,氨基酸L-天冬氨酸发挥以下作用:(1)假定递质;(2)蛋白质前体;(3)嘧啶和嘌呤碱基生物合成的原子供体;(4)能量代谢的燃料。星形胶质细胞在大脑中主导天冬氨酸的清除,在培养中它们摄取天冬氨酸并迅速将其代谢。在大脑中,只有星形胶质细胞被证明表达从头合成嘧啶的酶。为了更详细地了解天冬氨酸产生并随后由培养的星形胶质细胞释放的代谢物谱,对暴露于星形胶质细胞培养物的补充了[U-(13)C]天冬氨酸的孵育培养基进行了(13)C-核磁共振分析。结果表明,星形胶质细胞很容易代谢天冬氨酸,并将乳酸、谷氨酰胺、柠檬酸和丙氨酸的(13)C-同位素异构体释放到其培养基中。尽管星形胶质细胞中存在嘧啶生物合成的两种串联酶及其mRNA,但在培养基中未检测到嘧啶核苷酸相关的杂环化合物,如二氢乳清酸和乳清酸。

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