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Assessment of DNA damage in sperm after repeated non-invasive sampling in zebrafish Danio rerio

Authors

  • H. C. Reinardy,

    1. School of Biomedical and Biological Sciences, University of Plymouth, Devon, U.K.
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    • Present address: Bermuda Institute for Ocean Sciences, 17 Ferry Reach, St. George's, GE 01, Bermuda.

  • E. Skippins,

    1. School of Biomedical and Biological Sciences, University of Plymouth, Devon, U.K.
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  • T. B. Henry,

    Corresponding author
    1. Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN, U.S.A.
    2. Department of Forestry Wildlife and Fisheries, University of Tennessee, Knoxville, TN, U.S.A.
    • School of Biomedical and Biological Sciences, University of Plymouth, Devon, U.K.
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  • A. N. Jha

    1. School of Biomedical and Biological Sciences, University of Plymouth, Devon, U.K.
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Author to whom correspondence should be addressed. Tel.: +44 1752 232 940; email: Ted.Henry@ plymouth.ac.uk

Abstract

Repeated non-invasive sampling of zebrafish Danio rerio sperm was conducted, sperm counts were obtained and a method for measurement of DNA damage in sperm was developed and validated (single-cell gel electrophoresis, comet, assay). DNA damage in sperm increased with concentration of hydrogen peroxide (H2O2, 0–200 µM), and in vitro exposure of sperm to 200 µM H2O2 produced 88·7 ± 3·9% tail DNA compared to unexposed controls [12 ± 0·7% tail DNA (mean ± s.e., n = 3)]. Frequency of sperm sampling (sampled every 2, 4 or 7 days) did not affect DNA damage in sperm, but sperm counts decreased 57 and 22% for fish sampled every 2 or 4 days, respectively.

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