Detection of subcellular nitric oxide in mitochondria using a pyrylium probe: assays in cell cultures and peripheral blood
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Other documents of the author: Muñoz Resta, Ignacio; Bedrina Broch, Begoña; Martínez-Planes, Elena; Minguela, Alfredo; Galindo, Francisco
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comunitat-uji-handle3:10234/8639
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Title
Detection of subcellular nitric oxide in mitochondria using a pyrylium probe: assays in cell cultures and peripheral bloodAuthor (s)
Date
2021-11-25Publisher
Royal Society of ChemistryISSN
2050-750X; 2050-7518Bibliographic citation
RESTA, Ignacio Muñoz, et al. Detection of subcellular nitric oxide in mitochondria using a pyrylium probe: assays in cell cultures and peripheral blood. Journal of Materials Chemistry B, 2021, vol. 9, no 48, p. 9885-9892.Type
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Abstract
Fluorescent probes for the detection of intracellular nitric oxide (NO) are abundant, but those targeted to the mitochondria are scarce. Among those molecules targeting mitochondrial NO (mNO), the majority use a ... [+]
Fluorescent probes for the detection of intracellular nitric oxide (NO) are abundant, but those targeted to the mitochondria are scarce. Among those molecules targeting mitochondrial NO (mNO), the majority use a triphenylphosphonium (TPP) cation as a vector to reach such organelles. Here we describe a simple molecule (mtNOpy) based on the pyrylium structure, made in a few synthetic steps, capable of detecting selectively NO (aerated medium) over other reactive species. The calculated detection limit for mtNOpy is 88 nM. The main novelty of this probe is that it has a simple molecular architecture and can act both as a fluorogenic and as a mitochondriotropic agent, without using TPP. mtNOpy has been tested in two different scenarios: (a) in a controlled environment of cell line cultures (human colon carcinoma HT-29 cells and mouse macrophage RAW 264.7 cells), using confocal laser scanning microscopy, and (b) on a much more complex sample of peripheral blood, using flow cytometry. In the first context, mtNOpy has been found to be responsive (turn-on fluorescence) to exogenous and endogenous NO stimuli (via SNAP donor and LPS stimulation, respectively). In the second area, mtNOpy has been able to discriminate between NO-generating phagocytes (neutrophils and monocytes) from other leukocytes (NK, B and T cells). [-]
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J. Mater. Chem. B, 2021,9, 9885-9892Funder Name
Generalitat Valenciana | Santiago Grisolía
Project code
AICO/2020/322 | GRISOLIAP/2018/071
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This journal is © The Royal Society of Chemistry 2021
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