TY - JOUR
T1 - NOX5 in human spermatozoa
T2 - Expression, function, and regulation
AU - Musset, Boris
AU - Clark, Robert A.
AU - DeCoursey, Thomas E.
AU - Petheo, Gabor L.
AU - Geiszt, Miklos
AU - Chen, Yumin
AU - Cornell, John E.
AU - Eddy, Carlton A.
AU - Brzyski, Robert G.
AU - El Jamali, Amina
N1 - Copyright:
Copyright 2012 Elsevier B.V., All rights reserved.
PY - 2012/3/16
Y1 - 2012/3/16
N2 - Physiological and pathological processes in spermatozoa involve the production of reactive oxygen species (ROS), but the identity of the ROS-producing enzyme system(s) remains a matter of speculation. We provide the first evidence that NOX5 NADPH oxidase is expressed and functions in human spermatozoa. Immunofluorescence microscopy detected NOX5 protein in both the flagella/neck region and the acrosome. Functionally, spermatozoa exposed to calcium ionophore, phorbol ester, or H 2O 2 exhibited superoxide anion production, which was blocked by addition of superoxide dismutase, a Ca 2+ chelator, or inhibitors of either flavoprotein oxidases (diphenylene iododonium) or NOX enzymes (GKT136901). Consistent with our previous overexpression studies, we found that H 2O 2- induced superoxide production by primary sperm cells was mediated by the non-receptor tyrosine kinase c-Abl. Moreover, the H V1 proton channel, which was recently implicated in spermatozoa motility, was required for optimal superoxide production by spermatozoa. Immunoprecipitation experiments suggested an interaction among NOX5, c-Abl, and H V1. H 2O 2 treatment increased the proportion of motile sperm in a NOX5-dependent manner. Statistical analyses showed a pH-dependent correlation between superoxide production and enhanced sperm motility. Collectively, our findings show that NOX5 is a major source of ROS in human spermatozoa and indicate a role for NOX5-dependent ROS generation in human spermatozoa motility.
AB - Physiological and pathological processes in spermatozoa involve the production of reactive oxygen species (ROS), but the identity of the ROS-producing enzyme system(s) remains a matter of speculation. We provide the first evidence that NOX5 NADPH oxidase is expressed and functions in human spermatozoa. Immunofluorescence microscopy detected NOX5 protein in both the flagella/neck region and the acrosome. Functionally, spermatozoa exposed to calcium ionophore, phorbol ester, or H 2O 2 exhibited superoxide anion production, which was blocked by addition of superoxide dismutase, a Ca 2+ chelator, or inhibitors of either flavoprotein oxidases (diphenylene iododonium) or NOX enzymes (GKT136901). Consistent with our previous overexpression studies, we found that H 2O 2- induced superoxide production by primary sperm cells was mediated by the non-receptor tyrosine kinase c-Abl. Moreover, the H V1 proton channel, which was recently implicated in spermatozoa motility, was required for optimal superoxide production by spermatozoa. Immunoprecipitation experiments suggested an interaction among NOX5, c-Abl, and H V1. H 2O 2 treatment increased the proportion of motile sperm in a NOX5-dependent manner. Statistical analyses showed a pH-dependent correlation between superoxide production and enhanced sperm motility. Collectively, our findings show that NOX5 is a major source of ROS in human spermatozoa and indicate a role for NOX5-dependent ROS generation in human spermatozoa motility.
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U2 - 10.1074/jbc.M111.314955
DO - 10.1074/jbc.M111.314955
M3 - Article
C2 - 22291013
AN - SCOPUS:84863368859
VL - 287
SP - 9376
EP - 9388
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
SN - 0021-9258
IS - 12
ER -