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Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism.
Free Radic Biol Med 1999; 26(7-8):869-80FR

Abstract

Reactive oxygen species (ROS) inhibit sperm movement and have been implicated in male infertility. In this study, we determined the effects of specific ROS produced by activated leukocytes on human spermatozoa and investigated their metabolic site of action. We used chemiluminescence and electron paramagnetic resonance (EPR) to characterize the ROS generated by both blood and seminal leukocytes. We also determined the effects of these ROS on sperm energy metabolism using biochemical analyses and flow cytometry. Both blood and seminal leukocytes produced the same characteristic ROS which were determined to be hydrogen peroxide (H2O2) and superoxide radicals (O2*-). EPR using the spin trapping technique indicated that superoxide radical-dependent hydroxyl radicals (HO.) were also generated. ROS generated by PMA-stimulated blood leukocytes (2-5 x 10(6)/ml) caused inhibition of sperm movement in 2 h (p < .01). Using the hypoxanthine/ xanthine oxidase (0.5 U/ml) system to generate ROS, we determined that spermatozoa ATP levels, after ROS treatment, were reduced approximately eight-fold in 30 min (0.10 x 10(10) moles/10(6) sperm cells) compared to control (0.84 X 10(-10) moles/10(6) sperm cells) (p < .01). Sperm ATP reduction paralleled the inhibition of sperm forward progression. Neither superoxide dismutase (100 U/ml) nor dimethyl sulfoxide (100 mM) reversed these effects; however, protection was observed with catalase (4 X 10(3) U/ml). Flow cytometric analyses of sperm treated with various doses of H2O2 (0.3 mM-20.0 mM) showed a dose-dependent decrease in sperm mitochondrial membrane potential (MMP); however, at low concentrations of H2O2, sperm MMP was not significantly inhibited. Also, sperm MMP uncoupling with CCClP had no effect on either sperm ATP levels or forward progression. These results indicate that H2O2 is the toxic ROS produced by activated leukocytes causing the inhibition of both sperm movement and ATP production. O2*- and HO. do not play a significant role in these processes. Low concentrations of H2O2 causing complete inhibition of sperm movement and ATP levels inhibit sperm energy metabolism at a site independent of mitochondrial oxidative phosphorylation.

Authors+Show Affiliations

Department of Urology, Tulane University Medical Center, New Orleans, LA, USA.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

10232830

Citation

Armstrong, J S., et al. "Characterization of Reactive Oxygen Species Induced Effects On Human Spermatozoa Movement and Energy Metabolism." Free Radical Biology & Medicine, vol. 26, no. 7-8, 1999, pp. 869-80.
Armstrong JS, Rajasekaran M, Chamulitrat W, et al. Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radic Biol Med. 1999;26(7-8):869-80.
Armstrong, J. S., Rajasekaran, M., Chamulitrat, W., Gatti, P., Hellstrom, W. J., & Sikka, S. C. (1999). Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. Free Radical Biology & Medicine, 26(7-8), pp. 869-80.
Armstrong JS, et al. Characterization of Reactive Oxygen Species Induced Effects On Human Spermatozoa Movement and Energy Metabolism. Free Radic Biol Med. 1999;26(7-8):869-80. PubMed PMID: 10232830.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism. AU - Armstrong,J S, AU - Rajasekaran,M, AU - Chamulitrat,W, AU - Gatti,P, AU - Hellstrom,W J, AU - Sikka,S C, PY - 1999/5/8/pubmed PY - 1999/5/8/medline PY - 1999/5/8/entrez SP - 869 EP - 80 JF - Free radical biology & medicine JO - Free Radic. Biol. Med. VL - 26 IS - 7-8 N2 - Reactive oxygen species (ROS) inhibit sperm movement and have been implicated in male infertility. In this study, we determined the effects of specific ROS produced by activated leukocytes on human spermatozoa and investigated their metabolic site of action. We used chemiluminescence and electron paramagnetic resonance (EPR) to characterize the ROS generated by both blood and seminal leukocytes. We also determined the effects of these ROS on sperm energy metabolism using biochemical analyses and flow cytometry. Both blood and seminal leukocytes produced the same characteristic ROS which were determined to be hydrogen peroxide (H2O2) and superoxide radicals (O2*-). EPR using the spin trapping technique indicated that superoxide radical-dependent hydroxyl radicals (HO.) were also generated. ROS generated by PMA-stimulated blood leukocytes (2-5 x 10(6)/ml) caused inhibition of sperm movement in 2 h (p < .01). Using the hypoxanthine/ xanthine oxidase (0.5 U/ml) system to generate ROS, we determined that spermatozoa ATP levels, after ROS treatment, were reduced approximately eight-fold in 30 min (0.10 x 10(10) moles/10(6) sperm cells) compared to control (0.84 X 10(-10) moles/10(6) sperm cells) (p < .01). Sperm ATP reduction paralleled the inhibition of sperm forward progression. Neither superoxide dismutase (100 U/ml) nor dimethyl sulfoxide (100 mM) reversed these effects; however, protection was observed with catalase (4 X 10(3) U/ml). Flow cytometric analyses of sperm treated with various doses of H2O2 (0.3 mM-20.0 mM) showed a dose-dependent decrease in sperm mitochondrial membrane potential (MMP); however, at low concentrations of H2O2, sperm MMP was not significantly inhibited. Also, sperm MMP uncoupling with CCClP had no effect on either sperm ATP levels or forward progression. These results indicate that H2O2 is the toxic ROS produced by activated leukocytes causing the inhibition of both sperm movement and ATP production. O2*- and HO. do not play a significant role in these processes. Low concentrations of H2O2 causing complete inhibition of sperm movement and ATP levels inhibit sperm energy metabolism at a site independent of mitochondrial oxidative phosphorylation. SN - 0891-5849 UR - https://www.unboundmedicine.com/medline/citation/10232830/Characterization_of_reactive_oxygen_species_induced_effects_on_human_spermatozoa_movement_and_energy_metabolism_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0891-5849(98)00275-5 DB - PRIME DP - Unbound Medicine ER -