Fluorescence-Based Tool To Detect Endogenous Peroxynitrite in M1-Polarized Murine J774.2 Macrophages.

Clicks: 351
ID: 93794
2018
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Abstract
Oxidative stress and inflammation are intrinsically linked to each other. In addition, they are implicated in the evolution and progression of noncommunicable diseases (NCDs). Large amounts of reactive oxygen species (ROS) are generated as part of the immune response toward NCDs. Among all of the ROS species, peroxynitrite (ONOO) has the shortest half-life with <20 ms under typical physiological conditions. Hence, detecting ONOO and studying its generation in vitro allows for a better understanding of inflammatory processes. We demonstrate that peroxyresorufin-1 (PR1) is a selective and sensitive ONOO fluorescence-based sensor in J774.2 macrophages. PR1 was able to detect changes in ONOO production upon investigation of different factors: enhanced generation of ONOO through LPS and IFN-γ as well as diminished ONOO production with the introduction of superoxide scavengers and nitric oxide synthase inhibitors. Our study validates PR1 as an effective tool for the detection of ONOO in J774.2 murine macrophages and should allow for further elucidation of ROS biology and chemistry.
Reference Key
weber2018fluorescencebasedanalytical Use this key to autocite in the manuscript while using SciMatic Manuscript Manager or Thesis Manager
Authors Weber, Maria;Mackenzie, Amanda B;Bull, Steven D;James, Tony D;
Journal Analytical chemistry
Year 2018
DOI
10.1021/acs.analchem.8b03035
URL
Keywords
polarization light-field reflectance separation single-shot

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