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. 2018 Sep 21;6:270. doi: 10.3389/fpubh.2018.00270

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Copyright © 2018 Houston, Nixon, King, Aitken and De Iuliis.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The effects of RF-EMR on production of reactive oxygen species and lipid peroxidation in mature spermatozoa. Spermatozoa were isolated from the cauda epididymis and exposed to RF-EMR of an intensity of 0.15 W/kg. (A) Luminol-peroxidase chemiluminescence was used to assess ROS production following 4 h of exposure. (B) The lipid peroxidation status of these spermatozoa was evaluated with the BODIPY C11 probe via flow cytometry. (C) The profile of 4HNE alkylated-proteins in RF-EMR exposed spermatozoa was assessed via immunoblotting, with hexokinase expression featuring as a loading control. Three replicates were performed for both control and RF-EMR treated sperm protein extracts. (D) The corresponding intensity of all 4HNE labeled protein bands extracted from untreated control, and RF-EMR exposed spermatozoa, was determined by densitometric analysis of pixel intensity. Densitometry was performed on the principal bands from 70 to 40 kDa, relative to the 100 kDa hexokinase band presented below the 4HNE immunoblot (full blot shown in Supplementary Figure 1F).