EPR quantification of vascular nitric oxide production in genetically modified mouse models

被引:27
作者
Khoo, JP
Alp, NJ
Bendall, JK
Kawashima, S
Yokoyama, M
Zhang, YH
Casadei, B
Channon, KM [1 ]
机构
[1] Univ Oxford, John Radcliffe Hosp, Dept Cardiovasc Med, Oxford, England
[2] Kobe Univ, Sch Med, Dept Internal Med 1, Kobe, Hyogo 650, Japan
来源
NITRIC OXIDE-BIOLOGY AND CHEMISTRY | 2004年 / 10卷 / 03期
基金
英国惠康基金;
关键词
nitric oxide; measurement; EPR; colloid iron (II) diethyldithiocarbamate; eNOS knockout; eNOS-Tg; transgenic; aorta; mouse;
D O I
10.1016/j.niox.2004.04.003
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
With increasing use of genetically modified mice to study endothelial nitric oxide (NO) biology, methods for reliable quantification of vascular NO production by mouse tissues are crucial. We describe a technique based on electron paramagnetic resonance (EPR) spectroscopy, using colloid iron (11) diethyldithiocarbamate [Fe(DETC)(2)], to trap NO. A signal was seen from C57BL16 mice aortas incubated with Fe(DETC)(2), that increased 4.7-fold on stimulation with calcium ionophore A23187 [3.45 +/- 0.13 vs 0.73 +/- 0.13 au (arbitrary units)]. The signal increased linearly with incubation time (r(2) = 0.93), but was abolished by addition of N-G-nitro-L-arginine methyl ester (L-NAME) or endothelial removal. Stimulated aortas from eNOS knockout mice had virtually undetectable signals (0.14 +/- 0.06 vs 3.17 +/- 0.21 an in littermate controls). However, the signal was doubled from mice with transgenic eNOS overexpression (7.17 +/- 0.76 vs 3.37 +/- 0.43 au in littermate controls). We conclude that EPR is a useful too] for direct NO quantification in mouse vessels. (C) 2004 Elsevier Inc. All rights reserved.
引用
收藏
页码:156 / 161
页数:6
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