Skip to main content
SpringerLink
Log in

Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols

  • Published:
Cell Biology and Toxicology Aims and scope Submit manuscript

Abstract

The suitability of ejaculated bovine spermatozoa as an in vitro model for the assessment of the cytotoxic potential of chemicals was evaluated using several endpoints: swimming activity, adenine nucleotide content, membrane integrity and oxygen consumption. A series of chlorophenols inhibited sperm motion (motility and velocity) in a concentration-dependent manner. This could be determined quantitatively and reproducibly by means of videomicrography and automatic computer image analysis. The sperm immobilizing potency increased with increasing chlorination and was positively correlated with lipophilicity. Concentrations which reduced the percentage of moving sperm to 50% of controls ranged from 43 µM for pentachlorophenol (PCP) to 1440 µM for 4-monochlorophenol (4-MCP). Determinations of adenine nucleotides and percentages of viable cells revealed qualitative differences between the action of PCP and the lower chlorinated phenols. While the latter decreased the total adenine nucleotide contents and the percentage of unstained cells in parallel to motion inhibition, no such changes occurred after exposure to immobilizing concentrations of PCP. Penta-, tetra- and trichlorinated phenols stimulated cellular respiration, indicating their uncoupling activity, at concentrations lower than those necessary for motion inhibition. The results indicate that bovine spermatozoa may become a useful in vitro model for the toxicological evaluation of chemicals providing quantitative as well as qualitative data.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

PCP:

pentachlorophenol

2,3,4,5-TCP:

2,3,4,5-tetrachlorophenol

2,4,5-TCP:

2,4,5trichlorophenol

2,4-DCP:

2,4-dichlorophenol

4-MCP:

4-monochlorophenol

References

  • AITKEN, R.J., BEST, F.S.H. and RICHARDSON, D.W. (1982). The correlates of fertilizing capacity in normal fertile men. Fertil. Steril.38:68–76.

    Google Scholar 

  • ARNOLD, W.M., ZIMMERMANN, U., PAULI, W., BENZING, M., NIEHRS, C. and AHLERS, J. (1988). The comparative influence of substituted phenols (especially chlorophenols) on yeast cells assayed by electro-rotation and other methods. Biochim. Biophys. Acta942:83–95.

    Google Scholar 

  • BABICH, H. and BORENFREUND, E. (1987).In vitro cytotoxicity of organic pollutants to bluegill sunfish (BF-2) cells. Environm. Res.42:229–237.

    Google Scholar 

  • BENOIT-GUYOD, J.L., ANDRE, C., TAILLANDER, G., ROCHAT, JR. and BOUCHERLE, A. (1984). Toxicity and QSAR of chlorophenols on Lebistes reticulatus. Ecotox. Environm. Safety8:227–235.

    Google Scholar 

  • DEVILLERS, J. and CHAMBON, P. (1986). Acute toxicity and QSAR of chlorophenols on Daphnia magna. Bull. Environm. Contain. Toxicol.37:599–605.

    Google Scholar 

  • DUXBURY, C.L. and THOMPSON, J.E. (1987). Pentachlorophenol alters the molecular organization of membranes in mammalian cells. Arch. Environm. Contain. Toxicol.16:367–373.

    Google Scholar 

  • EKWALL, B., SELLING, J. and JOHNELS, D. (1987). Toxicity of chlorophenols to HeLa cells as measured in the MIT-24 system. ATLA14:178–181.

    Google Scholar 

  • ELIASSON, R. and TREICHL, L. (1971). Supravital staining of human spermatozoa. Fertil. Steril.22:134–137.

    Google Scholar 

  • FARQUHARSON, M.E., GAGE, J.C. and NORTHOVER, J. (1958). The biological action of chlorophenols. Br. J. Pharmacol.13:20–24.

    Google Scholar 

  • HALANGK, W. and BOHNENSACK, R. (1981). Besonderheiten der energiebereitstellenden Reaktionen in Bullenspermien und ihre Beziehung zur Vitalität. Arch. Exper. Vet. Med. 35:855–866.

    Google Scholar 

  • HALANGK, W., BOHNENSACK, R., FRANK, K. and KUNZ, W. (1985). Effect of various substrates on mitochondrial and cellular energy state of intact spermatozoa. Biomed. Biochim. Acta44:411–420.

    Google Scholar 

  • HONG, C.Y., CHAPUT DE SAINTONGE, D.M. and TURNER, P. (1981). A simple method to measure drug effects on human sperm motility. Br. J. Clin Pharmac.11:385–387.

    Google Scholar 

  • HONG, C.Y., CHAPUT DE SAINTONGE, D.M. and TURNER, P. (1982). Effects of chlorpromazine and other drugs acting on the central nervous system on human sperm motility. Europ. J. Clin. Pharmacol.22:413–416.

    Google Scholar 

  • HONG, C.Y. (1985). Pharmacological studies on human sperm motility. Trends Pharmacol. Sci.6:62–63.

    Google Scholar 

  • KALBE, L. and RECKNAGEL, R.-D. (1985). Zytotoxikologische Untersuchung von Schadstoffen und Wasserblüten mit gefrierkonserviertem Bullensperma (Spermatest). Acta Hydrochim. Hydrobiol.13:185–194.

    Google Scholar 

  • KATZ, D.F. and OVERSTREET, J.W. (1981). Sperm motility assessment by videomicrography. Fertil. Steril.35:188–193.

    Google Scholar 

  • KNASIAK-PALUCH, D. (1980). Toxicity of plastic materials as evaluated by means of bull semen (preliminary report). Pol. J. Pharmacol. Pharm.32:109–114.

    Google Scholar 

  • KÖNEMANN, H. and MUSCH, A. (1981). Quantitative structure-activity relationships in fish toxicity studies. Part 2: The influence of pH on the QSAR of chlorophenols. Toxicology19:223–228.

    Google Scholar 

  • KRAAYENHOF, F. and VAN DAM, K. (1969). Interaction between uncouplers and substrates in rat liver mitochondria. Biochim. Biophys. Acta172:189–197.

    Google Scholar 

  • LIU, Y.T. and WARME, P.K. (1977). Computerized evaluation of sperm cell motility. Computers. Biomed. Res.10:127–138.

    Google Scholar 

  • LYSIKIEWICZ, A. and ENHORNING, G. (1983). Tricolor photography for assessment of spermatozoan motility. Am. J. Obstet. Gynecol.145:229–233.

    Google Scholar 

  • MAKLER, A. (1980). Use of the elaborated multiple exposure photography (MEP) method in routine sperm motility analysis and for research purposes. Fertil. Steril.33:160–166.

    Google Scholar 

  • MAYEVSKY, A., BAR-SAGIE, D. and BARTOOV, B. (1983). Sperm cell motility as a new experimental model for toxicological studies. Arch. Toxicol. Suppl.6:295–299.

    Google Scholar 

  • NARDONE, R. (1983). The LD50-test andin vitro toxicology strategies. Acta Pharmacol. Toxicol.52 (Suppl. 11):65–79.

    Google Scholar 

  • NENDZA, M. (1987). Toxizitätsbestimmungen von umweltrelevanten Chemikalien mit einem neuen Biotestsystem. Ermittlung physikochemischer Eigenschaften und Ableitung quantitativer Struktur-Toxizitäts-Beziehungen unter Anwendung der Hauptkomponenten-Analyse. Thesis, University of Kiel.

  • OVERSTREET, J.W., KATZ, D.F., HANSON, F.W. and FONSECA, J.R. (1979). A simple inexpensive method for objective assessment of human sperm movement characteristics. Fertil. Steril.31:162–172.

    Google Scholar 

  • PALUCH, D., BORZEMSKA-SZYMONOWICZ, M. and OLSZEWSKA-BLACH, Z. (1980).In vitro screening studies of the toxicological testing of synthetic biomaterials. Introduction and comparative evaluation of a new method of testing applying breeding bull semen. Polymers in Med.X:193–204.

    Google Scholar 

  • RAOOF, N.T., PEARSON, R.M. and TURNER, P. (1987). A modified transmembrane migration method for measuring the effect of drugs on sperm motility. B. J. Clin. Pharmacol.24:319–321.

    Google Scholar 

  • SAARIKOSKI, J. and VILUKSELA, M. (1982). Relation between physicochemical properties of phenols and their toxicity and accumulation in fish. Ecotox. Environm. Safety6:501–512.

    Google Scholar 

  • SCHULTZ, T.W., HOLCOMBE, G.W. and PHIPPS, G.L. (1986). Relationships of quantitative structure-activity to comparative toxicity of selected phenols in the Pimephales promelas and Tetrahymena pyriformis test systems. Ecotoxicol. Environm. Safety12:146–153.

    Google Scholar 

  • SEIBERT, H. (1988). Messung der Bewegungsaktivität der Spermatozoen von Mensch und Rind mit Hilfe von Videomikrographie und Computerbildanalyse. Fertilität4:215–218.

    Google Scholar 

  • SPIELMANN, H., JACOB-MÜLLER, U. and SCHULZ, P. (1980). Simple assay of 0.1–1.0 pmol ATP, ADP and AMP in single somatic cells using purified luciferin luciferase. Anal. Biochem.113:172–178.

    Google Scholar 

  • STOCKDALE, M. and SELWYN, M.J. (1971). Effects of ring substituents on the activity of phenols as inhibitors and uncouplers of mitochondrial respiration. Eur. J. Biochem.21:565–574.

    Google Scholar 

  • VAN DOP, C., HUTSON, S.M. and LARDY, H.A. (1977). Pyruvate metabolism in bovine epididymal spermatozoa. J. Biol. Chem.252:1303–1308.

    Google Scholar 

  • WEINBACH, E.D. and GARBUS, J. (1965). The interaction of uncoupling phenols with mitochondria and with mitochondrial protein. J. Biol. Chem.240:1811–1819.

    Google Scholar 

  • ZABOJSZCZ, W., ZALEWSKI, W. and KOZIOROWSKA, J. (1980). Comparison of three methods of toxicity evaluation of medical plastics. Pol. J. Pharmacol. Pharm.32:99–102.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Toxicology, University of Kiel, Kiel, Federal Republic of Germany

    Hasso Seibert, Marike Kolossa & Otmar Wassermann

Authors
  1. Hasso Seibert
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Marike Kolossa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Otmar Wassermann
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Seibert, H., Kolossa, M. & Wassermann, O. Bovine spermatozoa as anin vitro model for studies on the cytotoxicity of chemicals: effects of chlorophenols. Cell Biol Toxicol 5, 315–330 (1989). https://doi.org/10.1007/BF01795359

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01795359

Key words

Search

Navigation