Effects of High Ambient Temperature on Fish Sperm Plasma Membrane Integrity and Mitochondrial Activity — A Flow Cytometric Study

Abstract

Local extreme climatic conditions occurring as a result of global climate change may interfere with the reproduction of animals. In the present study fish spermatozoa were incubated at different temperatures (20, 25, 30 and 40 °C) for 10 and 30 minutes, respectively and plasma membrane integrity and mitochondrial membrane potential changes were evaluated with flow cytometry using SYBR-14/PI and Mitotracker Deep Red FM fluorescent dyes. No significant differences were found in plasma membrane integrity at either incubation temperatures or time points. Mitotracker Deep Red FM histogram profiles indicating mitochondrial activity showed significant (p < 0.001) alterations in all cases of higher (25, 30 and 40 °C) temperature treatments as compared to the samples incubated at 20 °C. Our results indicate that fish spermatozoa exposed to high temperatures suffer sublethal damage that cannot be detected with conventional, vital staining techniques.

References

  1. 1.

    Aitken, R. J., De Iuliis G. N., Finnie J. M., Hedges A., McLachlan R. I. (2010) Analysis of the relationships between oxidative stress, DNA damage and sperm vitality in a patient population: development of diagnostic criteria. Hum. Reprod. 25, 2415–2426.

    CAS  Article  Google Scholar 

  2. 2.

    Barth, A. D., Oko, R. (1989) Abnormal morphology of bovine spermatozoa. Iowa State University Press, Ames.

    Google Scholar 

  3. 3.

    Bradshaw, W. E., Holzapfel, C. M. (2006) Evolutionary response to rapid climate change. Science 312, 1477–1478.

    CAS  Article  Google Scholar 

  4. 4.

    Bronson, F. (2009) Climate change and seasonal reproduction in mammals. Philos. T. Roy. Soc. B. 364, 3331–3340.

    CAS  Article  Google Scholar 

  5. 5.

    Bronson, F. H. (1989) Mammalian reproductive biology. University of Chicago Press, Chicago.

    Google Scholar 

  6. 6.

    Cox, C., Reeder, J. E., Robinson, R. D., Suppes, S. B., Wheeless, L. L. (1988) Comparison of frequency distributions in flow cytometry. Cytometry 9, 291–298.

    CAS  Article  Google Scholar 

  7. 7.

    Cummins, J. (2009) 5-Sperm motility and energetics. In: Pitnick, T. R. B. J. H. (ed.) Sperm Biology Academic Press, London, pp. 185–206.

    Google Scholar 

  8. 8.

    Garner, D., Johnson, L., Yue, S., Roth, B., Haugland, R. (1994) Dual DNA staining assessment of bovine sperm viability using SYBR-14 and propidium iodide. J. Androl. 15, 620–629.

    CAS  PubMed  Google Scholar 

  9. 9.

    Guthrie, H., Welch, G., Theisen, D., Woods, L. (2011) Effects of hypothermic storage on intracellular calcium, reactive oxygen species formation, mitochondrial function, motility, and plasma membrane integrity in striped bass (Morone saxatilis) sperm. Theriogenology 75, 951–961.

    CAS  Article  Google Scholar 

  10. 10.

    Guthrie, H., Woods, L., Long, J., Welch, G. (2008) Effects of osmolality on inner mitochondrial transmembrane potential and ATP content in spermatozoa recovered from the testes of striped bass (Morone saxatilis). Theriogenology 69, 1007–1012.

    CAS  Article  Google Scholar 

  11. 11.

    Hagedorn, M., Ricker, J., McCarthy, M., Meyers, S., Tiersch, T., Varga, Z., Kleinhans, F. (2009) Biophysics of zebrafish (Danio rerio) sperm. Cryobiology 58, 12–19.

    CAS  Article  Google Scholar 

  12. 12.

    Hallap, T., Nagy, S., Jaakma, Ü., Johannisson, A., Rodriguez-Martinez, H. (2005) Mitochondrial activity of frozen-thawed spermatozoa assessed by MitoTracker Deep Red 633. Theriogenology 63, 2311–2322.

    CAS  Article  Google Scholar 

  13. 13.

    Horváth, Á., Martínez-Páramo, S., Kovács, Á. I., Urbányi, B., Herráez, P. (2010) Effect of ovarian fluid on the mobility of fresh and cryopreserved sperm of the common carp (Cyprinus carpio L.). Állattani Közl. 95, 25–33. (In Hungarian)

    Google Scholar 

  14. 14.

    Hossain, M. S., Johannisson, A., Wallgren, M., Nagy, S., Siqueira, A. P., Rodriguez-Martinez, H. (2011) Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art. Asian J. Androl. 13, 406.

    Article  Google Scholar 

  15. 15.

    Inaba, K. (2008) Molecular mechanisms of the activation of flagellar motility in sperm. In: Alavi, S. M. H., Cosson, J. J., Coward, K., Rafiee, G. (ed.) Fish spermatology. Alpha Science International Ltd,, Oxford, UK, pp. 267–280.

    Google Scholar 

  16. 16.

    Ingermann, R. L. (2008) Energy metabolism and respiration in fish spermatozoa. In: Alavi, S. M. H., Cosson, J. J., Coward, K., Rafiee, G. (ed.) Fish spermatology. Alpha Science International Ltd., Oxford, UK, pp. 215–240.

    Google Scholar 

  17. 17.

    Jamieson, B. G. M. (1991) Fish Evolution and Systematics: Evidence from Spermatozoa: with a Survey of Lophophorate, Echinoderm, and Protochordate Sperm and an Account of Gamete Cryopreservation. Cambridge University Press, Cambridge.

    Google Scholar 

  18. 18.

    Jonsson, B., Jonsson N. (2009) A review of the likely effects of climate change on anadromous Atlantic salmon Salmo salar and brown trout Salmo trutta, with particular reference to water temperature and flow. J. Fish Biol. 75, 2381–2447.

    CAS  Article  Google Scholar 

  19. 19.

    Nagy, S., Kakasi, B., Havasi, M., Németh, S., Pál, L., Bercsényi, M., Husvéth, F. (2013) Dynamic cellular changes during fish sperm activation as measured by flow cytometry. Diversification in Inland Finfish Aquaculture II, Vodnany, Czech Republic.

    Google Scholar 

  20. 20.

    Pizzo, P., Drago, I., Filadi, R., Pozzan, T. (2012) Mitochondrial Ca2+ homeostasis: mechanism, role, and tissue specificities. Pflug. Arch. Eur. J. Phy. 464, 3–17.

    CAS  Article  Google Scholar 

  21. 21.

    Sood, S., Malecki, I., Tawang, A., Martin, G. (2012) Survival of emu (Dromaius novaehollandiae) sperm preserved at subzero temperatures and different cryoprotectant concentrations. Theriogenology 78, 1557–1569.

    CAS  Article  Google Scholar 

  22. 22.

    West, J. (2003) Effects of heat-stress on production in dairy cattle. J. Dairy Sci. 86, 2131–2144.

    CAS  Article  Google Scholar 

  23. 23.

    Young, I. T. (1977) Proof without prejudice: use of the Kolmogorov–Smirnov test for the analysis of histograms from flow systems and other sources. J. Histochem. Cytochem. 25, 935–941.

    CAS  Article  Google Scholar 

  24. 24.

    Zieba, G., Fox, M. G., Copp, G. H. (2010) The effect of elevated temperature on spawning of introduced pumpkinseed Lepomis gibbosus in Europe. J. Fish Biol. 77, 1850–1855.

    Article  Google Scholar 

Download references

Author information

Affiliations

Authors

Corresponding author

Correspondence to Szabolcs Tamás Nagy.

Rights and permissions

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Nagy, S.T., Kakasi, B., Pál, L. et al. Effects of High Ambient Temperature on Fish Sperm Plasma Membrane Integrity and Mitochondrial Activity — A Flow Cytometric Study. BIOLOGIA FUTURA 67, 125–132 (2016). https://doi.org/10.1556/018.67.2016.2.1

Download citation

Keywords

  • Fish sperm
  • plasma membrane integrity
  • mitochondrial activity
  • high temperature
  • flow cytometry