title: Boronate probes for the detection of hydrogen peroxide release from human spermatozoa.
abstract: Human spermatozoa are compromised by production of reactive oxygen species (ROS), and detection of ROS in spermatozoa is important for the diagnosis of male infertility. The probes 2',7'-dichlorohydrofluorescein diacetate (DCFH), dihydroethidium (DHE), and MitoSOX red (MSR) are commonly used for detecting ROS by flow cytometry; however, these probes lack sensitivity to hydrogen peroxide (H2O2), which is particularly damaging to mammalian sperm cells. This study reports the synthesis and use of three aryl boronate probes, peroxyfluor-1 (PF1), carboxyperoxyfluor-1, and a novel probe, 2-(2-ethoxyethoxy)ethoxyperoxyfluor-1 (EEPF1), in human spermatozoa. PF1 and EEPF1 were effective at detecting H2O2 and peroxynitrite (ONOO(-)) produced by spermatozoa when stimulated with menadione or 4-hydroxynonenal. EEPF1 was more effective at detection of ROS in spermatozoa than DCFH, DHE, or MSR; furthermore it distinguished poorly motile sperm as shown by greater ROS production. EEPF1 should therefore have a significant role in the diagnosis of oxidative stress in male infertility, cryopreservation, age, lifestyle, and exposure to environmental toxicants.
authors: ARC Centre of Excellence for Nanoscale BioPhotonics, Institute for Photonics & Advanced Sensing, and School of Physical Sciences, The University of Adelaide, Adelaide, SA 5005, Australia.
authors: Malcolm S
authors: MS
authors: Purdey
authors: Discipline of Biological Sciences and Priority Research Centre in Reproductive Science, Faculty of Science and IT, University of Newcastle, Callaghan, NSW 2308, Australia.
authors: Haley S
authors: HS
authors: Connaughton
authors: Discipline of Biological Sciences and Priority Research Centre in Reproductive Science, Faculty of Science and IT, University of Newcastle, Callaghan, NSW 2308, Australia.
authors: Sara
authors: S
authors: Whiting
authors: ARC Centre of Excellence for Nanoscale BioPhotonics, Institute for Photonics & Advanced Sensing, and School of Physical Sciences, The University of Adelaide, Adelaide, SA 5005, Australia.
authors: Erik P
authors: EP
authors: Schartner
authors: ARC Centre of Excellence for Nanoscale BioPhotonics, Institute for Photonics & Advanced Sensing, and School of Physical Sciences, The University of Adelaide, Adelaide, SA 5005, Australia.
authors: Tanya M
authors: TM
authors: Monro
authors: ARC Centre of Excellence for Nanoscale BioPhotonics, The Robinson Research Institute, and School of Paediatrics and Reproductive Health, The University of Adelaide, Adelaide, SA 5005, Australia.
authors: Jeremy G
authors: JG
authors: Thompson
authors: Discipline of Biological Sciences and Priority Research Centre in Reproductive Science, Faculty of Science and IT, University of Newcastle, Callaghan, NSW 2308, Australia. Electronic address: john.aitken@newcastle.edu.au.
authors: R John
authors: RJ
authors: Aitken
authors: ARC Centre of Excellence for Nanoscale BioPhotonics, Institute for Photonics & Advanced Sensing, and School of Physical Sciences, The University of Adelaide, Adelaide, SA 5005, Australia. Electronic address: andrew.abell@adelaide.edu.au.
authors: Andrew D
authors: AD
authors: Abell
mesh_terms: D000447:Aldehydes / Q000494:pharmacology; D001897:Boronic Acids / Q000138:chemical synthesis / Q000737:chemistry*; D002478:Cells, Cultured; D004996:Ethidium / Q000031:analogs & derivatives / Q000737:chemistry; D005452:Fluoresceins / Q000138:chemical synthesis / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000138:chemical synthesis / Q000737:chemistry*; D006801:Humans; D006861:Hydrogen Peroxide / Q000032:analysis / Q000378:metabolism; D008297:Male; D015335:Molecular Probes / Q000138:chemical synthesis / Q000737:chemistry*; D009943:Organophosphorus Compounds / Q000737:chemistry; D030421:Peroxynitrous Acid / Q000032:analysis / Q000378:metabolism; D010617:Phenanthridines / Q000737:chemistry; D013081:Sperm Motility / Q000187:drug effects; D013094:Spermatozoa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D024483:Vitamin K 3 / Q000494:pharmacology
chemical_list: D000447:Aldehydes; D001897:Boronic Acids; D005452:Fluoresceins; D005456:Fluorescent Dyes; C000597839:MitoSox Red; D015335:Molecular Probes; D009943:Organophosphorus Compounds; D010617:Phenanthridines; C500013:peroxyfluor-1; C067883:dihydroethidium; D030421:Peroxynitrous Acid; C029569:diacetyldichlorofluorescein; D024483:Vitamin K 3; D006861:Hydrogen Peroxide; D004996:Ethidium; C027576:4-hydroxy-2-nonenal
keywords: Aryl boronate; Free radicals; Human spermatozoa; Hydrogen peroxide; Organic synthesis; Reactive oxygen species