Abstract
Due to the increasing resistance of microbial pathogens to the available drugs, the identification of new antimicrobial agents with a new mechanism of action is urgently needed. In this context, cationic antimicrobial peptides (AMPs) are considered promising candidates. Although there is evidence that, in contrast to conventional antibiotics, microbial membranes are the principal target of a large number of AMPs, thus making it difficult for the pathogen to acquire resistance, their mode(s) of action is not yet completely clear. Intense research is currently devoted to understand the effect(s) of AMPs on intact cells, either at sub-lethal or at lethal peptide concentrations, and fluorescence/electron microscopy techniques represent a valid tool to get insight into the damage caused by these molecules on the morphology and membrane structure of the target cell. We here present an overview of some microscopic methodologies to address this issue.
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References
Boman, H. G. (1995) Peptide antibiotics and their role in innate immunity. Annu. Rev. Immunol. 13, 61–92.
Hancock, R. E. and Sahl, H. G. (2006) Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nat. Biotechnol. 24, 1551–1557.
Zasloff, M. (2002) Antimicrobial peptides of multicellular organisms. Nature 415, 389–395.
Jenssen, H., Hamill, P., and Hancock, R. E. (2006) Peptide antimicrobial agents. Clin. Microbiol. Rev. 19, 491–511.
Nicolas, P. and Mor, A. (1995) Peptides as weapons against microorganisms in the chemical defense system of vertebrates. Annu. Rev. Microbiol. 49, 277–304.
Mookherjee, N. and Hancock, R. E. (2007) Cationic host defence peptides: innate immune regulatory peptides as a novel approach for treating infections. Cell. Mol. Life Sci. 64, 922–933.
Mangoni, M. L., Papo, N., Barra, D., Simmaco, M., Bozzi, A., Di Giulio, A., and Rinaldi, A. C. (2004) Effects of the antimicrobial peptide temporin L on cell morphology, membrane permeability and viability of Escherichia coli. Biochem. J. 380, 859–865.
Rodriguez, G. G., Phipps, D., Ishiguro, K., and Ridgway, H. F. (1992) Use of a fluorescent redox probe for direct visualization of actively respiring bacteria. Appl. Environ. Microbiol. 58, 1801–1808.
Severin, E., Stellmach, J., and Nachtigall, H. M. (1985) Fluorimetric assay of redox activity in cells. Anal. Chim. Acta 170, 341–346.
Nation, J. L. (1983) A new method using hexamethyldisilazane for preparation of soft insect tissues for scanning electron microscopy. Stain Technol. 58, 347–351.
Acknowledgments
We thank Prof. Donatella Barra for careful reading the manuscript. Work reported here has been supported in part by grants from the Italian Ministry of Education, University and Research and from the Università di Roma “La Sapienza”.
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Marcellini, L., Giammatteo, M., Aimola, P., Mangoni, M.L. (2010). Fluorescence and Electron Microscopy Methods for Exploring Antimicrobial Peptides Mode(s) of Action. In: Giuliani, A., Rinaldi, A. (eds) Antimicrobial Peptides. Methods in Molecular Biology, vol 618. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-594-1_16
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DOI: https://doi.org/10.1007/978-1-60761-594-1_16
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