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Appl Environ Microbiol. 2015 Jul;81(14):4728-35. doi: 10.1128/AEM.00890-15. Epub 2015 May 8.

Use of Hybridization Chain Reaction-Fluorescent In Situ Hybridization To Track Gene Expression by Both Partners during Initiation of Symbiosis.

Author information

1
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA.
2
Department of Medical Microbiology and Immunology, University of Wisconsin-Madison, Madison, Wisconsin, USA mjmcfallngai@wisc.edu egruby@wisc.edu.

Abstract

The establishment of a productive symbiosis between Euprymna scolopes, the Hawaiian bobtail squid, and its luminous bacterial symbiont, Vibrio fischeri, is mediated by transcriptional changes in both partners. A key challenge to unraveling the steps required to successfully initiate this and many other symbiotic associations is characterization of the timing and location of these changes. We report on the adaptation of hybridization chain reaction-fluorescent in situ hybridization (HCR-FISH) to simultaneously probe the spatiotemporal regulation of targeted genes in both E. scolopes and V. fischeri. This method revealed localized, transcriptionally coregulated epithelial cells within the light organ that responded directly to the presence of bacterial cells while, at the same time, provided a sensitive means to directly show regulated gene expression within the symbiont population. Thus, HCR-FISH provides a new approach for characterizing habitat transition in bacteria and for discovering host tissue responses to colonization.

PMID:
25956763
PMCID:
PMC4551195
DOI:
10.1128/AEM.00890-15
[Indexed for MEDLINE]
Free PMC Article

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