Please use this identifier to cite or link to this item: https://hdl.handle.net/10316/105804
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 
POCI-01-0145-FEDER-007440 
CENTRO-01-0145-FEDER-000012-HealthyAging2020 
POCI-01-0145-FEDER-007440 
UID/NEU/04539/2019 
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.
URI: https://hdl.handle.net/10316/105804
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

Files in This Item:
File Description SizeFormat
Fluorescent_probes_for_the_detection_of_reactive.5.pdf3.33 MBAdobe PDFView/Open
Show full item record

Google ScholarTM

Check

Altmetric

Altmetric


This item is licensed under a Creative Commons License Creative Commons