Characterization of a fluorescent probe for imaging nitric oxide

Yohannes T. Ghebremariam, Ngan F. Huang, Swetha Kambhampati, Katharina S. Volz, Gururaj G. Joshi, Eric V. Anslyn, John P. Cooke

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

Background: Nitric oxide (NO), a potent vasodilator and anti-atherogenic molecule, is synthesized in various cell types, including vascular endothelial cells (ECs). The biological importance of NO enforces the need to develop and characterize specific and sensitive probes. To date, several fluorophores, chromophores and colorimetric techniques have been developed to detect NO or its metabolites (NO2 and NO3) in biological fluids, viable cells or cell lysates. Methods: Recently, a novel probe (NO550) has been developed and reported to detect NO in solutions and in primary astrocytes and neuronal cells with a fluorescence signal arising from a nonfluorescent background. Results: Here, we report further characterization of this probe by optimizing conditions for the detection and imaging of NO products in primary vascular ECs, fibroblasts, and embryonic stem cell-and induced pluripotent stem cell-derived ECs in the absence and presence of pharmacological agents that modulate NO levels. In addition, we studied the stability of this probe in cells over time and evaluated its compartmentalization in reference to organelle-labeling dyes. Finally, we synthesized an inherently fluorescent diazo ring compound (AZO550) that is expected to form when the nonfluorescent NO550 reacts with cellular NO, and compared its cellular distribution with that of NO550. Conclusion: NO 550 is a promising agent for imaging NO at baseline and in response to pharmacological agents that modulate its levels.

Original languageEnglish (US)
Pages (from-to)68-79
Number of pages12
JournalJournal of Vascular Research
Volume51
Issue number1
DOIs
StatePublished - Jan 2014

Keywords

  • Asymmetric dimethylarginine
  • Endothelial cells
  • Fluorescent NO probe
  • Nitric oxide
  • Nitric oxide imaging
  • Nitric oxide synthase

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

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