Isolation and properties of lead-resistant variants of rat glioma cells

  • Natalia Dolzhanskaya
  • Ekaterima Gomcharova
  • Toby G. Rossmam
Original Articles

Abstract

Glial cells are thought to protect neurons from heavy-metal toxicity. To gain a better understanding of mechanisms of protection against lead compounds, a number of lead-resistant C6 rat glioma cell sublines have been isolated. After 8 mo of growth in the absence of lead nitrate, three sublines still maintain their lead-resistant phenotype. None of the lead-resistant sublines are cross-resistant to Cd(II) or Ni(II), but all are cross-resistant (in varying degrees) to Hg(II), As(III), Sb(III), and Sn(II), and one is resistant to trimethyl tin. No inducible lead resistance is seen in any glioma line. One subline has been used to create cell-cell hybrids with wild-type cells. The hybrids exhibit dominance of the lead-resistant phenotype. To identify and analyze altered gene expression at the mRNA level in the lead-resistant sublines, the differential display technique was used. Numerous differences are seen between amplified fragments from wild-type and lead-resistant cells. Candidate clones are now being analyzed to confirm the differential expression and to isolate cDNAs that confer lead resistance.

Index entries

Lead arsenic cadmium tin nickel mercury antimony glioma cells 

References

  1. 1.
    C. R. Angle, Childhood lead poisoning and its treatment,Annu. Rev. Pharmacol. Toxicol. 33, 409–434 (1993).PubMedCrossRefGoogle Scholar
  2. 2.
    T. J. Simons, Lead-calcium interactions in cellular lead toxicity,NeuroToxicology 14, 77–86 (1993).PubMedGoogle Scholar
  3. 3.
    J. Markovac and G. W. Goldstein, Picomolar concentrations of lead stimulate brain protein kinase C,Nature 334, 71–73 (1988).PubMedCrossRefGoogle Scholar
  4. 4.
    J. T. Naarala, Lead amplifies glutamate-induced oxidative stress,Free Radical Biol. Med. 195, 689–693 (1995).CrossRefGoogle Scholar
  5. 5.
    P. Benda, J. Lightbody, G. Sato, L. Levine, and W. Sweet, Differentiated rat glial cell strain in tissue culture,Science 161, 370–371 (1968).PubMedCrossRefGoogle Scholar
  6. 6.
    C. G. Naus, K. Elisevich, D. Zhu, D. J. Belliveau, and R. F Del Maestro,In vivo growth of C6 glioma cells transfected with connexin 43 cDNA,Cancer Res. 52, 4208–4213 (1992).PubMedGoogle Scholar
  7. 7.
    E. Tiffany-Castiglioni, D. M. Garcia, J-N. Wu, J. Zmudzki, and G. R. Bratton, Effects of lead on viability and intracellular metal content of C6 rat glioma cells,J. Toxicol. Environ. Health,23, 267–279 (1988).PubMedCrossRefGoogle Scholar
  8. 8.
    E. Tiffany-Castiglioni, E. M. Sierra, J-N. Wu, and T. K. Rowles, Lead toxicity in neuroglia,NeuroToxicology 10, 417–44 (1989).PubMedGoogle Scholar
  9. 9.
    D. Holtzman, J. E. Olson, C. DeVries, and K. Bensch, Lead toxicity in primary cultured cerebral astrocytes and cerebellar granular neurons,Toxicol. Appl. Pharmacol. 89, 211–225 (1987).PubMedCrossRefGoogle Scholar
  10. 10.
    M. Aschner, N. B. Eberle, S. Goderie, and K. H. Kimelberg, Methylmercury uptake in rat primary astrocyte cultures: The role of the neutral amino acid transport system,Brain Res. 521, 221–228 (1990).PubMedCrossRefGoogle Scholar
  11. 11.
    A. M. Marini, J. P. Schwartz, and I. J. Kopin, The neurotoxicity of l-methyl-4-phenylpuridinium in cultured cerebellar granule cells,J. Neurosci. 9, 3665–3672 (1989).PubMedGoogle Scholar
  12. 12.
    P. Liang and A. B. Pardee, Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction,Science 257, 967–971 (1992).PubMedCrossRefGoogle Scholar
  13. 13.
    Z. Wang and T. G. Rossman, Stable and inducible arsenite resistance in Chinese hamster cells,Toxicol. Appl. Pharmacol. 118, 80–86 (1993).PubMedCrossRefGoogle Scholar
  14. 14.
    D. Gartside, B. Weiss, and H. L. Evans, Alkylmercurial encephalopathy in the monkey (Saimiri sciureus andMacaca arctoides): A histopathologic and autoradiographic study,Acta Neuropathol. 32, 61–74 (1975).CrossRefGoogle Scholar
  15. 15.
    B. Hitzfeld, F. Planas-Bohne, and D. Taylor, The effect of lead on protein and DNA metabolism of normal and lead-adapted rat kidney cells in culture,Biol. Trace Element Res. 21, 87–95 (1989).CrossRefGoogle Scholar
  16. 16.
    Y. Skreb, V. Habazin-Novak, and N. Hors, The rate of DNA synthesis in Hela cells during combined long-term and acute exposures to lead,Toxicology 19, 1–10 (1981).PubMedCrossRefGoogle Scholar
  17. 17.
    R. A. Goyer, P. May, M. M. Cats, and M. R. Krigman, Lead and protein content of isolated intranuclear inclusion bodies from kidneys of lead-poisoned rats,Lab. Invest. 22, 245–251 (1970).PubMedGoogle Scholar
  18. 18.
    B. A. Fowler, M. W. Kahng, D. R. Smith, E. A. Conner, and N. K. Laughlin, Implications of lead binding proteins for risk assessment of lead exposure,J. Expos. Anal. Environ. Epidemiol. 3, 441–448 (1993).Google Scholar
  19. 19.
    P. Mistry, G. Lucier, and B. Fowler, A High-affinity lead binding proteins in rat kidney cytosol mediate cell-free nuclear translocation of lead,J. Pharmacol. Exp. Ther. 232, 462 (1985).PubMedGoogle Scholar
  20. 20.
    Z. Wang, S. Dey, B. P. Rosen, and T. G. Rossman, Efflux mediated resistance to arsenicals in arsenic resistant and -hypersensitive Chinese hamster cells,Toxicol. Appl. Pharmacol. 137, 112–119 (1996).PubMedCrossRefGoogle Scholar
  21. 21.
    B. J. Aungst and H. L. Fung, Kinetic characterization of in vitro lead transport across the rat small intestine: Mechanism of intestinal lead transport,Toxicol. Appl. Pharmacol. 61, 39–47 (1981).PubMedCrossRefGoogle Scholar
  22. 22.
    J. L Tomsig and J. B. Suszkiw, Permeation of Pb2+ through calcium channels: fura-2 measurements of voltageand dihydropyridine-sensitive Pb2+ entry in isolated bovine chromaffin cells,Biochim. Biophys. Acta 1069, 197–200 (1991).PubMedCrossRefGoogle Scholar
  23. 23.
    G. Audesirk and T. Audesirk, The effects of inorganic lead on voltage-sensitive calcium channels differ among cell types and among channel subtypes,Neurotoxicology 14, 259–265 (1993).PubMedGoogle Scholar
  24. 24.
    Y. C. Qian, E. Tiffany-Castiglioni, and E. D. Harris, Copper transport and kinetics in cultured C6 rat glioma cells,Am. J. Physiol, Cell Physiol. 38, C892-C898 (1995).Google Scholar

Copyright information

© Humana Press Inc. 1998

Authors and Affiliations

  • Natalia Dolzhanskaya
    • 1
  • Ekaterima Gomcharova
    • 1
  • Toby G. Rossmam
    • 1
  1. 1.Nelson Institute of Environmental MedicineNew York University Medical CenterNew York

Personalised recommendations