Oxygen Consumption and Antioxidant Status of Plant Cells Cultured with Oxygenated Perfluorocarbon

  • Kenneth C. Lowe
  • Julie Wardrop
  • Paul Anthony
  • J. Brian Power
  • Michael R. Davey
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 540)

Abstract

Inert, respiratory gas-dissolving perfluorochemical (PFC) liquids can be used to regulate the supply of oxygen and carbon dioxide in both prokaryotic and eukaryotic (including human) cells in culture. However, the underlying biochemical changes in such cells, especially in relation to oxygen-sensitive pathways, are poorly understood. PFCs have been exploited as oxygen carriers to enhance mitosis during the culture of isolated protoplasts (naked cells from which the cell walls have been removed) of several plant species (Lowe et al., 1998), with subsequent increases in biomass from protoplast-derived cells. Whilst oxygen is vital in aerobic systems, its reduction may generate highly reactive oxygen species, such as superoxide (O2 -), which can have deleterious effects on cellular metabolism. Interestingly, there have been few studies on changes in cellular metabolism or on the activities of enzymes responsible for removal of reactive oxygen species in cells during culture with PFCs. Aerobic organisms have evolved enzyme systems for scavenging oxygen radicals, thus protecting against oxidative damage. One such group of enzymes, the superoxide dismutases (SOD), react with superoxide anions to produce hydrogen peroxide. The latter, in turn, is converted by catalases (CAT) to H2O and oxygen. This study has evaluated the beneficial effects of PFC-facilitated oxygen enhancement on changes in (1) cellular SOD (EC 1.15.1.1) and CAT (EC 1.11.1.6) activities, (2) the rate of oxygen consumption, as assessed by a Clark-type oxygen microelectrode, and (3) mitochondrial membrane potential (MMP) assessed using Rhodamine 123 fluorescence.

Keywords

Mitochondrial Membrane Potential Oxygen Carrier Mitotic Division Scavenge Oxygen Radical Cellular Oxygen Consumption 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. Aebi, H., 1984, Catalase in vitro. Methods Enzymol. 105: 121 - 126.CrossRefGoogle Scholar
  2. Beauchamp, C.O., and Fridovich, I., 1971, Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal. Biochem. 44: 276 - 287.CrossRefGoogle Scholar
  3. Branca, D., Chiarelli, S.M., Vincenti, E., Tortorella, C., and Scutari, G., 1994, Alteration of mitochondrial bioenergetics due to intravenous injection of a perfluorocarbon emulsion. Experientia 50: 660 - 663.PubMedCrossRefGoogle Scholar
  4. Frearson, E.M., Power, J.B., and Cocking, E.C., 1973, The isolation, culture and regeneration of Petunia leaf protoplasts. Dev. Biol. 33: 130 - 137.PubMedCrossRefGoogle Scholar
  5. Gilmour, D.M., Golds, T.J., and Davey, M.R., 1989, Medicago protoplasts: fusion, culture and plant regeneration, in: Biotechnology in Agriculture and Forestry, Plant Protoplasts and Genetic Engineering 1, Vol. 8, Y.P.S. Bajaj, ed., Springer-Verlag, Heidelberg, pp. 370 - 388.CrossRefGoogle Scholar
  6. Kao, K.N., and Michayluk, M.R., 1975, Nutritional requirements for growth of Vicia hajastana cells and protoplasts at a very low population density in liquid media. Planta 126: 105 - 110.CrossRefGoogle Scholar
  7. Lowe, K.C., Davey, M.R., and Power, J.B., 1998, Perfluorochemicals: their applications and benefits to cell culture. Trends Biotechnol. 16: 272 - 277.PubMedCrossRefGoogle Scholar
  8. Lowe, K.C., Davey, M.R., Power, J.B., and Mulligan, B.J., 1993, Surfactant supplements in plant culture systems. Agro-food-Ind. Hi-Tech. 4: 9 - 13.Google Scholar
  9. McCord, J.M., and Fridovich, I., 1969, Superoxide dismutase: an enzymatic function for erythocuprein (Hemocuprein). Biol. Chem. 244: 6049 - 6055.Google Scholar
  10. Porro, D., Smeraldi, C., Martegani, E., Ranzi, B.M., and Alberghina, L., 1994, Flow cytometric determination of the respiratory activity in growing Saccharomyces cerevisiae populations. Biotechnol. Prog. 10: 193197.Google Scholar
  11. Power, J.B., Davey, M.R., McLellan, M., and Wilson, D., 1990, Isolation, culture and fusion of protoplasts — 2. Fusion of protoplasts. Biotechnol. Educ. 1: 115 - 124.Google Scholar
  12. Snedecor, G.W., and Cochran, W.G., 1989, Statistical Methods, 8th edn., Iowa State College Press, Ames.Google Scholar
  13. Sureda, F.X., Escubedo, E., Gabriel, C., Comas, J., Camarasa, J., and Camins, A., 1997, Mitochondria) membrane potential measurement in rat cerebellar neurons by flow cytometry. Cytometry 28: 74 - 80.PubMedCrossRefGoogle Scholar
  14. Wardrop, J., Edwards, C.M., Lowe, K.C., Davey, M.R., and Power, J.B., 1997, Changes in cell biochemistry in response to culture of protoplasts with oxygenated perfluorocarbon. Art. Cells, Blood Subs, Immob. Biotechnol. 25: 585 - 589.Google Scholar
  15. Wardrop, J., Lowe, K.C., Power, J.B., and Davey, M.R., 1996, Perfluorochemicals and plant biotechnology: an improved protocol for protoplast culture and plant regeneration in rice (Oryza sativa L.). J. Biotechnol. 50: 47 - 54.CrossRefGoogle Scholar
  16. Widholm, J., 1972, The use of FDA and phenosafranine for determining viability of cultured plant cells. Stain Technol. 47: 186 - 194.Google Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Kenneth C. Lowe
    • 1
  • Julie Wardrop
    • 1
  • Paul Anthony
    • 2
  • J. Brian Power
    • 2
  • Michael R. Davey
    • 2
  1. 1.School of Life & Environmental SciencesUniversity of NottinghamNottinghamUK
  2. 2.School of BiosciencesUniversity of NottinghamLoughboroughUK

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