Definitive evidence for the nonmitochondrial production of superoxide anion by human spermatozoa

The Journal of Clinical Endocrinology and Metabolism
Geoffry N De IuliisR John Aitken

Abstract

Oxidative stress in the male germ line has been associated with poor fertility, impaired embryonic development, miscarriage, and childhood disease. Such stress is known to be associated with the peroxidation of unsaturated fatty acids in the sperm plasma membrane and oxidative DNA damage to both the nuclear and mitochondrial genomes. However, the source of the free radicals responsible for such damage is still unresolved. The objective of this study was to chemically validate the use of dihydroethidium (DHE) as a probe for detecting the generation of superoxide anion by human spermatozoa and to examine the relationship between this activity and defective sperm function. DHE and SYTOX green were used in conjunction with flow cytometry and HPLC to investigate superoxide generation by human spermatozoa. Cause and effect relationships were established using menadione to artificially drive superoxide production by these cells. HPLC, mass spectrometry, nuclear magnetic resonance (NMR) spectroscopy, and spectrofluorometry were used to demonstrate that human spermatozoa generate the superoxide-specific product, 2-hydroxyethidium, from DHE. Spontaneous superoxide production by human spermatozoa was found to originate from a nonmitochond...Continue Reading

Citations

Jun 21, 2007·Reproductive Biomedicine Online·R John Aitken, Geoffry N De Iuliis
Aug 4, 2009·Molecular Human Reproduction·R J Aitken, G N De Iuliis
Oct 23, 2009·Biology of Reproduction·Megan J McCarthyStuart A Meyers
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Jul 19, 2011·Experimental & Molecular Medicine·Sung Chun ChoSang Chul Park
Dec 20, 2011·Asian Journal of Andrology·Daniel R Franken, Sergio Oehninger
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Apr 20, 2007·Biology of Reproduction·D Claire WathesR John Aitken
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Jul 28, 2017·Molecular Reproduction and Development·Robert J Aitken
May 4, 2018·Andrology·S V A C SamarasingheR J Aitken

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