Tags

Type your tag names separated by a space and hit enter

Characterization of reactive oxygen species induced effects on human spermatozoa movement and energy metabolism.

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.

Links

  • Publisher Full Text
  • Authors+Show Affiliations

    ,

    Department of Urology, Tulane University Medical Center, New Orleans, LA, USA.

    , , , ,

    Source

    Free radical biology & medicine 26:7-8 1999 Apr pg 869-80

    MeSH

    Adenosine Triphosphate
    Carbonyl Cyanide m-Chlorophenyl Hydrazone
    Catalase
    Dimethyl Sulfoxide
    Electron Spin Resonance Spectroscopy
    Energy Metabolism
    Humans
    Hydrogen Peroxide
    Hypoxanthine
    In Vitro Techniques
    Leukocytes
    Luminescent Measurements
    Male
    Reactive Oxygen Species
    Sperm Motility
    Spermatozoa
    Superoxide Dismutase
    Superoxides
    Tetradecanoylphorbol Acetate
    Xanthine Oxidase

    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 -