Quantification of superoxide radical formation in intact vascular tissue using aCypridinaLuciferin analog as an alternative to lucigenin
MP Skatchkov, D Sperling, U Hink, E Änggard… - Biochemical and …, 1998 - Elsevier
MP Skatchkov, D Sperling, U Hink, E Änggard, T Münzel
Biochemical and biophysical research communications, 1998•ElsevierLucigenin has been frequently used for the chemiluminescent detection of superoxide (• O−
2) in intact tissue. More recent studies, however, revealed that lucigenin per se causes
formation of• O− 2raising doubt about this probe to detect reliably• O− 2. We therefore tested
a more recently described chemiluminescence probe (2-methyl-6-phenyl-3, 7-
dihydroimidazol [1, 2-α] pyrazine-3-one (CLA)) to estimate the ability of vascular tissue to
generate• O− 2as an alternative to lucigenin. In a cell free system as well as in vascular …
2) in intact tissue. More recent studies, however, revealed that lucigenin per se causes
formation of• O− 2raising doubt about this probe to detect reliably• O− 2. We therefore tested
a more recently described chemiluminescence probe (2-methyl-6-phenyl-3, 7-
dihydroimidazol [1, 2-α] pyrazine-3-one (CLA)) to estimate the ability of vascular tissue to
generate• O− 2as an alternative to lucigenin. In a cell free system as well as in vascular …
Lucigenin has been frequently used for the chemiluminescent detection of superoxide (•O−2) in intact tissue. More recent studies, however, revealed that lucigenin per se causes formation of•O−2raising doubt about this probe to detect reliably•O−2. We therefore tested a more recently described chemiluminescence probe (2-methyl-6-phenyl-3,7-dihydroimidazol[1,2-α]pyrazine-3-one (CLA)) to estimate the ability of vascular tissue to generate•O−2as an alternative to lucigenin. In a cell free system as well as in vascular tissue, CLA-enhanced chemiluminescence was dose dependently inhibited by superoxide dismutase (SOD), vitamin C and sodium nitroprusside (SNP). Electron spin resonance studies revealed that lucigenin (250μM) but not CLA (1 μM) caused extra•O−2production in vascular tissue. Stimulation of vessels with NADH (200μM) increased CLA enhanced chemiluminescence, which was inhibited by low concentrations of superoxide dismutase (20U/ml). Endothelial removal as well as the nitric oxidase-synthase inhibitor increased CLA chemiluminescence in vessels. We conclude that CLA is a sensitive and specific chemiluminescence probe to detect•O−2production in intact vascular tissue.
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