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Title: Fluorescent probes for the detection of reactive oxygen species in human spermatozoa
Authors: Escada-Rebelo, Sara 
Mora, Francisca 
Sousa, Ana 
Almeida Santos, Teresa 
Paiva, Artur 
Ramalho-Santos, João 
Keywords: flow cytometry; fluorescent probes; human spermatozoa; oxidative stress; reactive oxygen species
Issue Date: 2020
Publisher: Wolters Kluwer Health
Project: PD/BD/128237/2016 – PhD Programme in Experimental Biology and Biomedicine 
Serial title, monograph or event: Asian Journal of Andrology
Volume: 22
Issue: 5
Abstract: Reactive oxygen species (ROS) production is a by-product of mitochondrial activity and is necessary for the acquisition of the capacitated state, a requirement for functional spermatozoa. However, an increase in oxidative stress, due to an abnormal production of ROS, has been shown to be related to loss of sperm function, highlighting the importance of an accurate detection of sperm ROS, given the specific nature of this cell. In this work, we tested a variety of commercially available fluorescent probes to detect ROS and reactive nitrogen species (RNS) in human sperm, to define their specificity. Using both flow cytometry (FC) and fluorescence microscopy (FM), we confirmed that MitoSOX™ Red and dihydroethidium (DHE) detect superoxide anion (as determined using antimycin A as a positive control), while DAF-2A detects reactive nitrogen species (namely, nitric oxide). For the first time, we also report that RedoxSensor™ Red CC-1, CellROX® Orange Reagent, and MitoPY1 seem to be mostly sensitive to hydrogen peroxide, but not superoxide. Furthermore, mean fluorescence intensity (and not percentage of labeled cells) is the main parameter that can be reproducibly monitored using this type of methodology.
ISSN: 1008-682X
DOI: 10.4103/aja.aja_132_19
Rights: openAccess
Appears in Collections:I&D CNC - Artigos em Revistas Internacionais
IIIUC - Artigos em Revistas Internacionais
I&D ICBR - Artigos em Revistas Internacionais

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