Correlative analyses of nitric oxide generation rates and nitric oxide synthase levels in individual cells using a modular cell-retaining device
Autor: | Elena Afrimzon, Yishay Tauber, Naomi Zurgil, Yana Shafran, Maria Sobolev, Mordechai Deutsch, Asher Shainberg |
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Rok vydání: | 2012 |
Předmět: |
Cell
Nitric Oxide Synthase Type II Nanotechnology Nitric Oxide Analytical Chemistry Nitric oxide Promonocyte Cell Line chemistry.chemical_compound Immune system medicine Humans Fluorescent Dyes chemistry.chemical_classification Microscopy Confocal biology Lysophosphatidylcholines Glutathione Immunohistochemistry Cell biology Up-Regulation Nitric oxide synthase Enzyme medicine.anatomical_structure chemistry biology.protein Fluorescein Intracellular |
Zdroj: | Analytical chemistry. 84(17) |
ISSN: | 1520-6882 |
Popis: | Nitric oxide (NO) is recognized as one of the major immune system agents involved in the pathogenesis and control of various diseases that may benefit from novel drug development, by exploiting NO signaling pathways and targets. This calls for detection of both intracellular levels of NO and expression of its synthesizing enzymes (NOS) in individual, intact, living cells. Such measurements are challenging, however, due to short half-life, low and fluctuating concentrations of NO, cellular heterogeneity, and inability to trace the same cells over time. The current study presents a device and methodology for correlative analysis of NO generation rates and NOS levels in the same individual cells, utilizing fluorescent imaging followed by immunohistochemistry (IHC). U937 promonocyte cell populations demonstrated significant heterogeneity in their baseline levels, in NO-generation kinetics, and in their response rates to stimuli. Individual cell analysis exposed cell subgroups which showed enhanced NO production upon stimulation, concomitantly with significant up-regulation of inducible NOS (iNOS) levels. Exogenous NO modulated the expression of iNOS in nondifferentiated cells within 1 h, in a dose-dependent manner, while treatment with lysophosphatidylcholine (LPC) enhanced the expression of iNOS, demonstrating a nondependence on NO production. |
Databáze: | OpenAIRE |
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