Detection of nitric oxide formation in primary neural cells and tissues
Document Type
Article
Abstract
Nitric oxide (NO) is a free radical molecule with a short half-life (<5 s). Because its synthesis from L-arginine by constitutive NO synthase (NOS) is low in many cell types, including neurons and endothelial cells, direct detection of NO in biological systems is a difficult task. During pathological conditions in the CNS, the inducible form of NOS (iNOS or NOS2) is expressed in activated astrocytes and microglial cells and can result in higher levels of NO. However, it may still be difficult to detect NO in these cell types using typical spectrophotometric methods. Of particular note, NO is readily oxidized to nitrite and nitrate (relatively stable products) in cells and medium, which can be measured as a valid indicator of NO synthesis. The conversion of NO to peroxynitrite leads to the formation of stable protein adducts that can be detected by immunohistochemical or immunofluorescence methods. Additionally, intracellular levels of NO can be detected in real time using fluorescence imaging and NO-specific, cell permeable indicator dyes.
Medical Subject Headings
Animals; Astrocytes (metabolism); Cells, Cultured; Chromatography, High Pressure Liquid (methods); Mice; Microscopy, Fluorescence; Nitric Oxide (chemistry, metabolism); Peroxynitrous Acid (metabolism); Primary Cell Culture; Single-Cell Analysis (methods); Tissue Culture Techniques
Publication Date
1-1-2011
Publication Title
Methods in molecular biology (Clifton, N.J.)
E-ISSN
1940-6029
Volume
758
First Page
267
Last Page
77
PubMed ID
21815072
Digital Object Identifier (DOI)
10.1007/978-1-61779-170-3_18
Recommended Citation
Tjalkens, Ronald B.; Carbone, David L.; and Wu, Guoyao, "Detection of nitric oxide formation in primary neural cells and tissues" (2011). Translational Neuroscience. 2513.
https://scholar.barrowneuro.org/neurobiology/2513