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Quorum Sensing pp 219-233 | Cite as

Linking Quorum Sensing Regulation and Biofilm Formation by Candida albicans

  • Aurélie Deveau
  • Deborah A. Hogan
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 692)

Abstract

Candida albicans biofilms are surface-associated, structured communities composed of yeast, hyphal, and pseudohyphal cells surrounded by an extracellular matrix. C. albicans biofilms often lead to life-threatening systemic infections and are particularly difficult to eradicate because of their high levels of resistance to antibiotics. Farnesol, an autoregulatory molecule secreted by C. albicans, inhibits hyphal growth and the expression of a number of morphology-specific genes that are necessary for robust biofilm formation. Many stages of biofilm development are impacted by farnesol including the adherence of cells to the substratum, the architecture of mature biofilms, and the dispersal of cells from biofilms. For these reasons, understanding the mechanisms of action of farnesol could lead to the development of new antifungal compounds that target C. albicans biofilm cells, perhaps rendering biofilms more sensitive to antibiotics. Here, we describe several methods for the analysis of the effects of farnesol on biofilm formation and function.

Key words

Farnesol Biofilm Candida albicans Morphology 

Notes

Acknowledgment

We thank Dr. Aaron Mitchell (Carnegre mellon University, PA, USA) for providing CSLM pictures. The data described in this chapter were generated in studies funded by NIH (K22 DE016542, D.A.H.).

References

  1. 1.
    Ramage, G., Saville, S. P., Thomas, D. P., and Lopez-Ribot, J. L. (2005) Candida biofilms: an update. Eukaryot Cell 4, 633–8.PubMedCrossRefGoogle Scholar
  2. 2.
    Chandra, J., Kuhn, D. M., Mukherjee, P. K., Hoyer, L. L., McCormick, T., and Ghannoum, M. A. (2001) Biofilm formation by the fungal pathogen Candida albicans: development, architecture, and drug resistance. J Bacteriol 183, 5385–94.PubMedCrossRefGoogle Scholar
  3. 3.
    Al-Fattani, M. A., and Douglas, L. J. (2006) Biofilm matrix of Candida albicans and Candida tropicalis: chemical composition and role in drug resistance. J Med Microbiol 55, 999–1008.PubMedCrossRefGoogle Scholar
  4. 4.
    Nobile, C. J., Nett, J. E., Hernday, A. D., Homann, O. R., Deneault, J. S., Nantel, A., Andes, D. R., Johnson, A. D., and Mitchell, A. P. (2009) Biofilm matrix regulation by Candida albicans Zap1. PLoS Biol 7, e1000133.PubMedCrossRefGoogle Scholar
  5. 5.
    Parsek, M. R., and Greenberg, E. P. (2005) Sociomicrobiology: the connections between quorum sensing and biofilms. Trends Microbiol 13, 27–33.PubMedCrossRefGoogle Scholar
  6. 6.
    Blankenship, J. R., and Mitchell, A. P. (2006) How to build a biofilm: a fungal perspective. Curr Opin Microbiol 9, 588–94.PubMedCrossRefGoogle Scholar
  7. 7.
    Hornby, J. M., Jensen, E. C., Lisec, A. D., Tasto, J. J., Jahnke, B., Shoemaker, R., Dussault, P., and Nickerson, K. W. (2001) Quorum sensing in the dimorphic fungus Candida albicans is mediated by farnesol. Appl Environ Microbiol 67, 2982–92.PubMedCrossRefGoogle Scholar
  8. 8.
    Ramage, G., Saville, S. P., Wickes, B. L., and Lopez-Ribot, J. L. (2002) Inhibition of Candida albicans biofilm formation by farnesol, a quorum-sensing molecule. Appl Environ Microbiol 68, 5459–63.PubMedCrossRefGoogle Scholar
  9. 9.
    Nickerson, K. W., Atkin, A. L., and Hornby, J. M. (2006) Quorum sensing in dimorphic fungi: farnesol and beyond. Appl Environ Microbiol 72, 3805–13.PubMedCrossRefGoogle Scholar
  10. 10.
    Westwater, C., Balish, E., and Schofield, D. A. (2005) Candida albicans-conditioned medium protects yeast cells from oxidative stress: a possible link between quorum sensing and oxidative stress resistance. Eukaryot Cell 4, 1654–61.PubMedCrossRefGoogle Scholar
  11. 11.
    Davis-Hanna, A., Piispanen, A. E., Stateva, L. I., and Hogan, D. A. (2008) Farnesol and dodecanol effects on the Candida albicans Ras1-cAMP signalling pathway and the regulation of morphogenesis. Mol Microbiol 67, 47–62.PubMedCrossRefGoogle Scholar
  12. 12.
    Shirtliff, M. E., Krom, B. P., Meijering, R. A., Peters, B. M., Zhu, J., Scheper, M. A., Harris, M. L., and Jabra-Rizk, M. A. (2009) Farnesol-induced apoptosis in Candida albicans. Antimicrob Agents Chemother 53, 2392–401.PubMedCrossRefGoogle Scholar
  13. 13.
    Chen, H., Fujita, M., Feng, Q., Clardy, J., and Fink, G. R. (2004) Tyrosol is a quorum-sensing molecule in Candida albicans. Proc Natl Acad Sci USA 101, 5048–52.PubMedCrossRefGoogle Scholar
  14. 14.
    Alem, M. A., Oteef, M. D., Flowers, T. H., and Douglas, L. J. (2006) Production of tyrosol by Candida albicans biofilms and its role in quorum sensing and biofilm development. Eukaryot Cell 5, 1770–9.PubMedCrossRefGoogle Scholar
  15. 15.
    Baillie, G. S., and Douglas, L. J. (1999) Role of dimorphism in the development of Candida albicans biofilms. J Med Microbiol 48, 671–9.PubMedCrossRefGoogle Scholar
  16. 16.
    Ramage, G., VandeWalle, K., Lopez-Ribot, J. L., and Wickes, B. L. (2002) The filamentation pathway controlled by the Efg1 regulator protein is required for normal biofilm formation and development in Candida albicans. FEMS Microbiol Lett 214, 95–100.PubMedCrossRefGoogle Scholar
  17. 17.
    Richard, M. L., Nobile, C. J., Bruno, V. M., and Mitchell, A. P. (2005) Candida albicans biofilm-defective mutants. Eukaryot Cell 4, 1493–502.PubMedCrossRefGoogle Scholar
  18. 18.
    Nobile, C. J., Andes, D. R., Nett, J. E., Smith, F. J., Yue, F., Phan, Q. T., Edwards, J. E., Filler, S. G., and Mitchell, A. P. (2006) Critical role of Bcr1-dependent adhesins in Candida albicans biofilm formation in vitro and in vivo. PLoS Pathog 2, e63.PubMedCrossRefGoogle Scholar
  19. 19.
    Harcus, D., Nantel, A., Marcil, A., Rigby, T., and Whiteway, M. (2004) Transcription profiling of cyclic AMP signaling in Candida albicans. Mol Biol Cell 15, 4490–9.PubMedCrossRefGoogle Scholar
  20. 20.
    Bahn, Y. S., Molenda, M., Staab, J. F., Lyman, C. A., Gordon, L. J., and Sundstrom, P. (2007) Genome-wide transcriptional profiling of the cyclic AMP-dependent signaling pathway during morphogenic transitions of Candida albicans. Eukaryot Cell 6, 2376–90.PubMedCrossRefGoogle Scholar
  21. 21.
    Smith, D. A., Nicholls, S., Morgan, B. A., Brown, A. J., and Quinn, J. (2004) A conserved stress-activated protein kinase regulates a core stress response in the human pathogen Candida albicans. Mol Biol Cell 15, 4179–90.PubMedCrossRefGoogle Scholar
  22. 22.
    Kruppa, M., Krom, B. P., Chauhan, N., Bambach, A. V., Cihlar, R. L., and Calderone, R. A. (2004) The two-component signal transduction protein Chk1p regulates quorum sensing in Candida albicans. Eukaryot Cell 3, 1062–5.PubMedCrossRefGoogle Scholar
  23. 23.
    Kebaara, B. W., Langford, M. L., Navarathna, D. H., Dumitru, R., Nickerson, K. W., and Atkin, A. L. (2008) Candida albicans Tup1 is involved in farnesol-mediated inhibition of filamentous-growth induction. Eukaryot Cell 7, 980–7.PubMedCrossRefGoogle Scholar
  24. 24.
    Navarathna, D. H., Hornby, J. M., Krishnan, N., Parkhurst, A., Duhamel, G. E., and Nickerson, K. W. (2007) Effect of farnesol on a mouse model of systemic candidiasis, determined by use of a DPP3 knockout mutant of Candida albicans. Infect Immun 75, 1609–18.PubMedCrossRefGoogle Scholar
  25. 25.
    Nett, J., and Andes, D. (2006) Candida albicans biofilm development, modeling a host–pathogen interaction. Curr Opin Microbiol 9, 340–5.PubMedCrossRefGoogle Scholar
  26. 26.
    Chandra, J., Mukherjee, P. K., and Ghannoum, M. A. (2008) In vitro growth and analysis of Candida biofilms. Nat Protoc 3, 1909–24.PubMedCrossRefGoogle Scholar
  27. 27.
    Heydorn, A., Nielsen, T. A., Hentzer, M., Sternberg, C., Givskov, M., Ersboll, B., and Molin, S. (2000) Quantification of biofilm structures by the novel computer program COMSTAT. Microbiology 146, 2395–2407.PubMedGoogle Scholar
  28. 28.
    Ramage, G., Vande Walle, K., Wickes, B. L., and Lopez-Ribot, J. L. (2001) Standardized method for in vitro antifungal susceptibility testing of Candida albicans biofilms. Antimicrob Agents Chemother 45, 2475–9.PubMedCrossRefGoogle Scholar
  29. 29.
    Pierce, C. G., Thomas, D. P., and Lopez-Ribot, J. L. (2009) Effect of tunicamycin on Candida albicans biofilm formation and maintenance. J Antimicrob Chemother 63, 473–9.PubMedCrossRefGoogle Scholar
  30. 30.
    Martins, M., Henriques, M., Azeredo, J., Rocha, S. M., Coimbra, M. A., and Oliveira, R. (2007) Morphogenesis control in Candida albicans and Candida dubliniensis through signaling molecules produced by planktonic and biofilm cells. Eukaryot Cell 6, 2429–36.PubMedCrossRefGoogle Scholar
  31. 31.
    Weber, K., Sohr, R., Schulz, B., Fleischhacker, M., and Ruhnke, M. (2008) Secretion of E,E-farnesol and biofilm formation in eight different Candida species. Antimicrob Agents Chemother 52, 1859–61.PubMedCrossRefGoogle Scholar
  32. 32.
    Krom, B. P., Cohen, J. B., McElhaney Feser, G. E., and Cihlar, R. L. (2007) Optimized candidal biofilm microtiter assay. J Microbiol Methods 68, 421–23.PubMedCrossRefGoogle Scholar
  33. 33.
    Chandra, J., Mukherjee, P. K., Leidich, S. D., Faddoul, F. F., Hoyer, L. L., Douglas, L. J., and Ghannoum, M. A. (2001) Antifungal resistance of candidal biofilms formed on denture acrylic in vitro. J Dent Res 80, 903–8.PubMedCrossRefGoogle Scholar
  34. 34.
    Uppuluri, P., Chaturvedi, A. K., and Lopez-Ribot, J. L. (2009) Design of a simple model of Candida albicans biofilms formed under conditions of flow: development, architecture, and drug resistance. Mycopathologia 168, 101–9.PubMedCrossRefGoogle Scholar
  35. 35.
    Nikawa, H., Jin, C., Hamada, T., Makihira, S., Kumagai, H., and Murata, H. (2000) Interactions between thermal cycled resilient denture lining materials, salivary and serum pellicles and Candida albicans in vitro. Part II. Effects on fungal colonization. J Oral Rehabil 27, 124–30.PubMedCrossRefGoogle Scholar
  36. 36.
    Peeters, E., Nelis, H. J., and Coenye, T. (2008) Comparison of multiple methods for quantification of microbial biofilms grown in microtiter plates. J Microbiol Methods 72, 157–65.PubMedCrossRefGoogle Scholar
  37. 37.
    Nobile, C. J., and Mitchell, A. P. (2005) Regulation of cell-surface genes and biofilm formation by the Candida albicans transcription factor Bcr1p. Curr Biol 15, 1150–5.PubMedCrossRefGoogle Scholar
  38. 38.
    Hogan, D. A., Vik, A., and Kolter, R. (2004) A Pseudomonas aeruginosa quorum-sensing molecule influences Candida albicans morphology. Mol Microbiol 54, 1212–23.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Aurélie Deveau
    • 1
  • Deborah A. Hogan
    • 1
  1. 1.Department of Microbiology and ImmunologyDartmouth Medical SchoolHanoverUSA

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