Abstract
The chemiluminescence method is the most commonly employed technique as a direct measurement of reactive oxygen species (ROS) generation by spermatozoa. This assay is capable of quantifying both intracellular and extracellular ROS. Moreover, the use of various probes allows for differentiation between superoxide and hydrogen peroxide production by spermatozoa. When the total antioxidant reserves are overwhelmed by excessive production of ROS, it results in oxidative stress. Therefore correct measurement of both ROS and total antioxidant capacity (TAC) is essential in the assessment of oxidative stress in sperm and semen. This chapter describes the methodological approach for measuring seminal oxidative stress through the use of chemiluminescence assay for accurate measurement of ROS and the colorimetric assay for measurement of TAC.
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References
Sharma RK et al (1999) The reactive oxygen species-total antioxidant capacity score is a new measure of oxidative stress to predict male infertility. Hum Reprod 14:2801–2807
Aitken RJ, Buckingham D (1992) Enhanced detection of reactive oxygen species produced by human spermatozoa with 7-dimethyl amino-naphthalin-1, 2-dicarbonic acid hydrazide. Int J Androl 15:211–219
Aitken RJ et al (1992) Reactive oxygen species and human spermatozoa: analysis of the cellular mechanisms involved in luminol- and lucigenin-dependent chemiluminescence. J Cell Physiol 151:466–477
Sharma RK, Agarwal A (1996) Role of reactive oxygen species in male infertility. Urology 48:835–850
Kobayashi H et al (2001) Quality control of reactive oxygen species measurement by luminol-dependent chemiluminescence assay. J Androl 22:568–574
Allamaneni SS et al (2005) Characterization of oxidative stress status by evaluation of reactive oxygen species levels in whole semen and isolated spermatozoa. Fertil Steril 83:800–803
Agarwal A et al (2004) Chemiluminescence technique for measuring reactive oxygen species. Reprod Biomed Online 9:466–468
Allamaneni SS et al (2005) Comparative study on density gradients and swim-up preparation techniques utilizing neat and cryopreserved spermatozoa. Asian J Androl 7:86–92
Saleh RA, Agarwal A (2002) Oxidative stress and male infertility: from research bench to clinical practice. J Androl 23:737–752
Saleh RA et al (2002) Leukocytospermia is associated with increased reactive oxygen species production by human spermatozoa. Fertil Steril 78:1215–1224
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Kashou, A.H., Sharma, R., Agarwal, A. (2013). Assessment of Oxidative Stress in Sperm and Semen. In: Carrell, D., Aston, K. (eds) Spermatogenesis. Methods in Molecular Biology, vol 927. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-038-0_30
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DOI: https://doi.org/10.1007/978-1-62703-038-0_30
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-037-3
Online ISBN: 978-1-62703-038-0
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