Fluorescent probes for the detection of reactive oxygen species in human spermatozoa

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...

Full description

Saved in:
Bibliographic Details
Published in:Asian journal of andrology 2020-09, Vol.22 (5), p.465-471
Main Authors: Escada-Rebelo, Sara, Mora, Francisca, Sousa, Ana, Almeida-Santos, Teresa, Paiva, Artur, Ramalho-Santos, João
Format: Article
Language:English
Subjects:
EDTA
flow cytometry
fluorescent probes
human spermatozoa
oxidative stress
reactive oxygen species
Investigations
Original
Oxidation
Reactive oxygen species
Sperm
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary: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
1745-7262
DOI:10.4103/aja.aja_132_19