Abstract
A large number of bacterial species move smoothly on solid surfaces in the absence of extracellular organelles. In the deltaproteobacterium Myxococcus xanthus, this surface motion, termed gliding motility, involves a novel macromolecular machinery Agl-Glt. During the motility process, the Agl-Glt system, an integral envelope protein complex, is assembled on the ventral side of the cell. Doing so, the complex couples surface adhesion to the activity of the Agl motility motor. On the cytosolic side, the Agl-Glt system is linked to the bacterial actin cytoskeleton MreB. It is proposed that motility is produced when surface immobilized Agl-Glt complexes produce traction on a rigid track, possibly the MreB cables. Testing this hypothesis directly requires both microfluidic techniques to perturb the motility process with inhibitors (i.e., A22, CCCP) and state-of-the-art microscopy techniques (i.e., TIRF and AFM). These approaches require a microscopy chamber where the cells glide in liquid on a non-agar substrate. Here, we describe a straightforward coating procedure to construct a chitosan-functionalized microfluidic chamber that fulfills these requirements. This set up circumvents all the disadvantages of traditional agar-based assays, providing new grounds for high-resolution experiments. We also describe simple image processing to maximize the quality of data representation. In theory, our procedure could be used for any bacterial system that adheres to chitosan.
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References
Shrout JD, Tolker-Nielsen T, Givskov M, Parsek MR (2011) The contribution of cell-cell signaling and motility to bacterial biofilm formation. MRS Bull 36:367–373
Curtis PD, Taylor RG, Welch RD, Shimkets LJ (2007) Spatial organization of Myxococcus xanthus during fruiting body formation. J Bacteriol 189:9126–9130
McBride MJ (2001) Bacterial gliding motility: multiple mechanisms for cell movement over surfaces. Annu Rev Microbiol 55:49–75
Kearns DB (2010) A field guide to bacterial swarming motility. Nat Rev Microbiol 8:634–644
Merz AJ, So M, Sheetz MP (2000) Pilus retraction powers bacterial twitching motility. Nature 407:98–102
Skerker JM, Berg HC (2001) Direct observation of extension and retraction of type IV pili. Proc Natl Acad Sci U S A 98:6901–6904
McBride MJ (2004) Cytophaga-flavobacterium gliding motility. J Mol Microbiol Biotechnol 7:63–71
Luciano J et al (2011) Emergence and modular evolution of a novel motility machinery in bacteria. PLoS Genet 7:e1002268
Nan B, Mauriello EMF, Sun I-H, Wong A, Zusman DR (2010) A multi-protein complex from Myxococcus xanthus required for Bacterial gliding motility. Mol Microbiol 76:1539–1554
Sun M, Wartel M, Cascales E, Shaevitz JW, Mignot T (2011) Motor-driven intracellular transport powers bacterial gliding motility. Proc Natl Acad Sci 108:7559–7564
Nan B et al (2011) Myxobacteria gliding motility requires cytoskeleton rotation powered by proton motive force. Proc Natl Acad Sci U S A 108:2498–2503
Mauriello EMF et al (2010) Bacterial motility complexes require the actin-like protein, MreB and the Ras homologue, MglA. EMBO J 29:315–326
Mignot T, Shaevitz JW, Hartzell PL, Zusman DR (2007) Evidence that focal adhesion complexes power bacterial gliding motility. Science 315:853–856
Ducret A et al (2009) A microscope automated fluidic system to study bacterial processes in real time. PLoS One 4:e7282
Berginski ME, Vitriol EA, Hahn KM, Gomez SM (2011) High-resolution quantification of focal adhesion spatiotemporal dynamics in Living cells. PLoS One 6:e22025
Ng JMK, Gitlin I, Stroock AD, Whitesides GM (2002) Components for integrated poly(dimethylsiloxane) microfluidic systems. Electrophoresis 23: 3461–3473.
Acknowledgments
We thank Yong Zhang for suggesting Chitosan as a potential motility substrate. AD was funded by an HFSP young investigator grant (HFSP—RGY0075/2008) to TM. This work was supported by a C-Nano PACA grant (AAP09) to OT and TM.
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Ducret, A., Théodoly, O., Mignot, T. (2013). Single Cell Microfluidic Studies of Bacterial Motility. In: Delcour, A. (eds) Bacterial Cell Surfaces. Methods in Molecular Biology, vol 966. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-245-2_6
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DOI: https://doi.org/10.1007/978-1-62703-245-2_6
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