Subcellular analysis of D-aspartate.
Subcellular analysis of D-aspartate.
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DOI：
10.1021/ac0511694
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发表时间：
2005-10
影响因子：
7.4
通讯作者：
Hai Miao;S. Rubakhin;J. Sweedler
中科院分区：
文献类型：
--
作者：
Hai Miao;S. Rubakhin;J. Sweedler
D-Aspartate (D-Asp) is an especially intriguing molecule found within neurons of the central nervous system of animals ranging from mollusks to vertebrates. It has a large variety of roles ascribed to it, including an involvement in cell-to-cell signaling. To determine the D-Asp content in cells and in subcellular domains, a laboratory-assembled capillary electrophoresis system with laser-induced fluorescence (LIF) detection has been used. The system allows chiral separations with sufficient sensitivity and selectivity to measure the D-Asp content in specific subregions of a single neuron, including neuronal processes. The method uses microvial sampling, analyte derivatization with naphthalene-2,3-dicarboxaldehyde, cyclodextrin-mediated micellar electrokinetic capillary chromatography, and sheath flow cell-based LIF detection. Manipulating neuronal processes is difficult as they often disintegrate during the transfer to the sampling vial. We describe a glycerol treatment that stabilizes cell morphology during sample preparation, thereby alleviating the deleterious effects of the high-salt extracellular matrix on the electrophoretic separation. D-Asp percentages in processes from identified neurons from Aplysia californica differ significantly depending on the cell studied. Subcellular analysis reveals more compounds in the cell body than in the processes.
