Capillary electrophoresis analysis of nitric oxide synthase related metabolites in single identified neurons
Capillary electrophoresis analysis of nitric oxide synthase related metabolites in single identified neurons
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DOI：
10.1021/ac9713013
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发表时间：
1998-06-01
影响因子：
7.4
通讯作者：
Sweedler, JV
中科院分区：
文献类型：
--
作者：
Floyd, PD;Moroz, LL;Sweedler, JV
L-瓜氨酸（Cit）及其代谢物的细胞内浓度与一氧化氮合酶（NOS）活性有关，NOS是一种在动物组织（包括神经系统）中产生细胞间信使NO的酶。本文描述了一种采用激光诱导荧光检测的毛细管电泳系统，并开发了监测已鉴定的海蛞蝓、侧鳃亚纲和加利福尼亚无鳃海蛞蝓神经元中L-精氨酸（Arg）、Cit和相关分子水平的方法。和L-精氨酸磷酸盐的范围从50阿莫尔到17 fmol（在所研究的神经元中为5 nM到17 μ M）;这些检测限显著低于代谢物的实际细胞内水平，允许直接测定单细胞。单个神经元中NOS代谢物的水平从假定的含NOS神经元中的6（Arg）和4 mM（Cit）变化到
Intracellular concentrations of L-citrulline (Cit) and its metabolites are related to nitric oxide synthase (NOS) activity, an enzyme producing the intercellular messenger NO in animal tissues including the nervous system. A capillary electrophoresis system using laser-induced fluorescence detection is described, and methods are developed to monitor the levels of L-arginine (Arg), Cit, and related molecules in identified neurons of the marine slugs, Pleurobranchaea californica and Aplysia californica, The limits of detection for Arg, Cit, L-argininosuccinate, L-ornithine, and L-arginine phosphate range from 50 amol to 17 fmol (5 nM to 17 mu M in the neurons under study); these detection limits are significant lower than actual intracellular levels of the metabolites, allowing the direct assay of single cells. The levels of NOS metabolites in individual neurons varied from 6 (Arg) and 4 mM (Cit) in putative NOS-containing neurons down to