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BIOLOGICAL SCIENCES / MICROBIOLOGY
In vivo imaging and genetic analysis link bacterial motility and symbiosis in the zebrafish gut

John F. Rawls^*,,, Michael A. Mahowald^*, Andrew L. Goodman^*, Chad M. Trent, and Jeffrey I. Gordon^*,

*Center for Genome Sciences, Washington University School of Medicine, St. Louis, MO 63108; and Department of Cell and Molecular Physiology, University of North Carolina, Chapel Hill, NC 27599

Contributed by Jeffrey I. Gordon, March 14, 2007 (received for review January 26, 2007)

Complex microbial communities reside within the intestines of humans and other vertebrates. Remarkably little is known about how these microbial consortia are established in various locations within the gut, how members of these consortia behave within their dynamic ecosystems, or what microbial factors mediate mutually beneficial host–microbial interactions. Using a gnotobiotic zebrafish–Pseudomonas aeruginosa model, we show that the transparency of this vertebrate species, coupled with methods for raising these animals under germ-free conditions can be used to monitor microbial movement and localization within the intestine in vivo and in real time. Germ-free zebrafish colonized with isogenic P. aeruginosa strains containing deletions of genes related to motility and pathogenesis revealed that loss of flagellar function results in attenuation of evolutionarily conserved host innate immune responses but not conserved nutrient responses. These results demonstrate the utility of gnotobiotic zebrafish in defining the behavior and localization of bacteria within the living vertebrate gut, identifying bacterial genes that affect these processes, and assessing the impact of these genes on host–microbial interactions.

Danio rerio | establishment of a gut microbiota | flagellar motility | host–microbial symbiosis and mutualism | Pseudomonas aeruginosa

Author contributions: J.F.R., M.A.M., and J.I.G. designed research; J.F.R., M.A.M., and C.M.T. performed research; J.F.R., M.A.M., A.L.G., and J.I.G. analyzed data; and J.F.R., M.A.M., A.L.G., and J.I.G. wrote the paper.

The authors declare no conflict of interest.

This article contains supporting information online at www.pnas.org/cgi/content/full/0702386104/DC1.

To whom correspondence may be addressed. E-mail: jfrawls{at}med.unc.edu or jgordon{at}wustl.edu

© 2007 by The National Academy of Sciences of the USA

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