Abstract
Purpose
To investigate the influence of sperm DNA integrity on the zona binding ability of mouse spermatozoa in relation to their sex chromosomal constitution.
Method(s)
In this prospective experimental study, the sperm DNA fragmentation was induced by exposing testicular area of Swiss Albino mice (Mus musculus) to different doses of γ-radiation (0, 2.5, 5.0 and 10.0 Gy). Sperm DNA fragmentation was quantified by single cell gel electrophoresis (comet assay). In vitro sperm zona binding assay was performed and the numbers of zona bound X and Y bearing spermatozoa were determined using fluorescence in situ hybridization (FISH).
Result(s)
The assessment of zona pellucida bound X and Y-bearing spermatozoa using fluorescence in situ hybridization has revealed a unique binding pattern. The number of zona bound Y-spermatozoa declined significantly (P < 0.01 to 0.0001) with increase in the DNA damage. The skewed binding pattern of X and Y-bearing sperm was strongly correlated with the extent of sperm DNA damage.
Conclusion(s)
The zona pellucida may have a role in preventing DNA damaged mouse sperm binding especially towards Y-bearing sperm. However, the exact mechanism behind this observation needs to be elucidated further.
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Acknowledgments
Part of this work was supported by Indian Council of Medical Research in the form of senior research fellowship to DU (IRISID No. 2006–01640). Authors are thankful to Radiation Oncology department for providing radiation facility.
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Differences in zona pellucida binding of mouse X and Y bearing spermatozoa are related to their DNA integrity.
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Supplementary Figure 1
Fluorescent in situ hybridization (FISH) of zona pellucida bound X-bearing (green) and Y-bearing (red) spermatozoa. (JPEG 22 kb)
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Kumar, D., Upadhya, D., Uppangala, S. et al. Nuclear DNA fragmentation negatively affects zona binding competence of Y bearing mouse spermatozoa. J Assist Reprod Genet 30, 1611–1615 (2013). https://doi.org/10.1007/s10815-013-0123-x
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DOI: https://doi.org/10.1007/s10815-013-0123-x