Establishment of a Sensitive Nitric Oxide Bioassay for Bioscreening Application

Abstract

Nitric oxide is a free-radical diatomic molecule and an important biologic messenger molecule involved in the regulation of diverse physiological and pathophysiological mechanisms in the cardiovascular system, the central and peripheral nervous systems and the immune system. The discovery of NO with these important physiological functions has led to the development of various analytical methods for its detection and quantification. The involvement of nitric oxide in a plethora of vital biological functions makes its detection and quantification of great interest. Among various methods of measurement of the production of nitric oxide in cellular systems, those based on the use of fluorogenic probes have been gaining popularity due to their simplicity and sensitivity. BODIPY has advantages of high extinction coefficients, high fluorescence quantum efficiency and stability to light. Moreover, its emission wavelength can be adjusted by modifying chemical groups. Our lab has been exploring functionalized DABs for labeling and detection of NO several years before. This study was aimed at an evaluation of the specificity of reaction of DAMBO-PH with NO, in particular at an assessment of its reactivity with nitric oxide and its bioscreening application to biological systems.