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D-serine uptake and release in PC-12 cells measured by chiral microchip electrophoresis-mass spectrometry.

Abstract：

:Previous work has established that D-serine (D-Ser) plays important roles in certain neurological processes. Study on its uptake/storage and release by neuronal cells is highly significant for elucidating relevant mechanisms. In this work, PC-12 cells were incubated with racemic Ser (100 μM each enantiomer). After incubation, both intra- and extracellular levels of D-Ser and L-Ser were quantified by chiral microchip electrophoresis with mass spectrometric detection. It was found the cells preferably took up D-Ser over L-Ser. After 120 min incubation, D-Ser percentage ([D-Ser]/([D-Ser] + [L-Ser]) in the culture media changed from 50% to 9% while inside the cells it increased from 13% to 67%. Small neutral amino acids such as threonine impaired D-Ser uptake. Ser release was studied by using PC-12 cells preloaded with D-Ser. KCl, Glu, and Gly evoked Ser release. Interestingly, while depolarization by KCl evoked release of Ser as a D-Ser/L-Ser mixture of 1:1 ratio, the stereoisomeric composition of Ser released due to Glu exposure varied with the exposure time, ranging from 73% D-Ser (i.e., [D-Ser] > [L-Ser]) at 2 min to 44% (i.e., [D-Ser] < [L-Ser]) at 14 min, clearly indicating a stereochemical preference for D-Ser in Ser release from neuronal cells evoked by Glu-receptor activation.

journal_name

ACS Chem Neurosci

journal_title

ACS chemical neuroscience

authors

Li X,McCullum C,Zhao S,Hu H,Liu YM

doi

10.1021/cn5003122

subject

Has Abstract

pub_date

2015-04-15 00:00:00

pages

582-7

issue

4

issn

1948-7193

journal_volume

6

pub_type

杂志文章

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