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Assessing Pseudomonas aeruginosa Autoinducer Effects on Mammalian Epithelial Cells
Department of Surgery, Texas Tech University Health Sciences Center, MS8312, 3601 4th Street, Lubbock, USA.
Department of Surgery, Texas Tech University Health Sciences Center, MS8312, 3601 4th Street, Lubbock, USA.
Department of Surgery, Texas Tech University Health Sciences Center, MS8312, 3601 4th Street, Lubbock, USA.
Linköping University, Department of Clinical and Experimental Medicine, Division of Microbiology and Molecular Medicine. Linköping University, Faculty of Medicine and Health Sciences.
2018 (English)In: Quorum Sensing: Methods and Protocols / [ed] Livia LeoniGiordano Rampioni, Humana Press, 2018, Vol. 1673, p. 213-225Chapter in book (Refereed)
Abstract [en]

The human mucosal environment in the gut is rich with interactions between microbiota and mammalian epithelia. Microbes such as the Gram-negative bacterium Pseudomonas aeruginosa may use quorum sensing to communicate with other microorganisms and mammalian cells to alter gene expression. Here, we present methodologies to evaluate the effects of P. aeruginosa N-(3-oxo-dodecanoyl)-L-homoserine lactone (3O-C12-HSL) on Caco-2 cell monolayers. First, we describe a method for assessing barrier function and permeability of epithelial cells when exposed to 3O-C12-HSL by measuring transepithelial electrical resistance (TER) and paracellular flow using fluorescently labeled dextran. Secondly, we detail methods to investigate the effect of 3O-C12-HSL on protein-protein interactions of epithelial junction proteins. Lastly, we will detail imaging techniques to visualize Caco-2 barrier disruption following exposure to 3O-C12-HSL through the use of confocal laser scanning microscopy (CLSM) and a super resolution technique, stimulated emission depletion (STED) microscopy, to achieve nanoscale visualization of Caco-2 monolayers.

Place, publisher, year, edition, pages
Humana Press, 2018. Vol. 1673, p. 213-225
Series
Methods in Molecular Biology, ISSN 1064-3745, E-ISSN 1940-6029 ; 1673
Keywords [en]
Acyl homoserine lactone; Cell junction; Host pathogen interaction; IQGAP1; Imaging; Paracellular permeability; Pseudomonas aeruginosa; Quorum sensing; Transepithelial electrical resistance
National Category
Cell Biology
Identifiers
URN: urn:nbn:se:liu:diva-152539DOI: 10.1007/978-1-4939-7309-5_17PubMedID: 29130176ISBN: 9781493973088 (print)ISBN: 9781493973095 (electronic)OAI: oai:DiVA.org:liu-152539DiVA, id: diva2:1299564
Available from: 2019-03-27 Created: 2019-03-27 Last updated: 2019-03-27Bibliographically approved

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Vikström, Elena

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