title: DHR123: an alternative probe for assessment of ROS in human spermatozoa.
abstract: The aim of this study was to assess the potential of dihydrorhodamine 123 (DHR123) to measure oxidative stress produced by human spermatozoa. The results were compared with 2', 7'-dichlorodihydrofluorescein diacetate (DCFH-DA) that is routinely used for assessment of H(2)O(2) produced by spermatozoa. Fluorescence intensity and percentage R123 and DCF positive sperm were measured by flow cytometry. The optimal condition for assessment of reactive oxygen species (ROS) produced by sperm with DHR123 was 0.05 µM for 1 million sperm per ml for 20 minutes. The results of ROS measurement by DHR123 showed a significant correlation (r= +0.818, P<0.001) with DCFH-DA staining. Immunofluorescence of sperm stained with DHR123 revealed ROS production in the sperm mid-piece. In addition a significant correlation was observed between oxidant production assessed by DHR123 and semen parameters. Therefore, DHR123 may be considered a suitable probe for estimating oxidants produced by human spermatozoa, and can present heterogeneity in oxidant production between different samples.
authors.affiliation: Department of Reproduction and Development, Reproductive Biomedicine Center, Royan Institute for Animal Biotechnology, ACECR, Isfahan, Iran.
authors.forename: Abbas
authors.initials: A
authors.lastname: Kiani-Esfahani
authors.forename: Marzeyeh
authors.initials: M
authors.lastname: Tavalaee
authors.forename: Mohammad R
authors.initials: MR
authors.lastname: Deemeh
authors.forename: Mohammad
authors.initials: M
authors.lastname: Hamiditabar
authors.forename: Mohammad H
authors.initials: MH
authors.lastname: Nasr-Esfahani
mesh_terms: D000328:Adult; D006801:Humans; D006861:Hydrogen Peroxide / Q000378:metabolism; D007248:Infertility, Male / Q000473:pathology; D008297:Male; D008875:Middle Aged; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism*; D012235:Rhodamines / Q000737:chemistry / Q000378:metabolism*; D013076:Sperm Count; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology* / Q000378:metabolism
chemical_list: D017382:Reactive Oxygen Species; D012235:Rhodamines; C058319:dihydrorhodamine 123; D006861:Hydrogen Peroxide