AI Article Synopsis

  • Current methods for studying microbial gene expression in infected tissues are limited to bulk analyses, which overlook the diversity of individual cells and the tissue's structure.
  • This study introduces a new technique, HCR RNA-FISH, that allows visualization and quantification of gene transcripts at a single-cell level in the infected tongues of mice.
  • By using this method, researchers observed significant variations between cells and unique patterns of mRNA expression related to oral candidiasis, enhancing our understanding of how fungal pathogens interact with host tissues.

Article Abstract

Unlabelled: Microbial gene expression measurements derived from infected organs are invaluable to understand pathogenesis. However, current methods are limited to "bulk" analyses that neglect microbial cell heterogeneity and the lesion's spatial architecture. Here, we report the use of hybridization chain reaction RNA fluorescence hybridization (HCR RNA-FISH) to visualize and quantify transcripts at single-cell resolution in tongues of infected mice. The method is compatible with fixed-frozen and formalin-fixed paraffin-embedded tissues. We document cell-to-cell variation and intriguing spatiotemporal expression patterns for mRNAs that encode products implicated in oral candidiasis. The approach provides a spatial dimension to gene expression analyses of host- interactions.

Importance: is a fungal pathobiont inhabiting multiple mucosal surfaces of the human body. Immunosuppression, antibiotic-induced microbial dysbiosis, or implanted medical devices can impair mucosal integrity enabling to overgrow and disseminate, causing either mucosal diseases such as oropharyngeal candidiasis or life-threatening systemic infections. Profiling fungal genes that are expressed in the infected mucosa or in any other infected organ is paramount to understand pathogenesis. Ideally, these transcript profiling measurements should reveal the expression of any gene at the single-cell level. The resolution typically achieved with current approaches, however, limits most gene expression measurements to cell population averages. The approach described in this report provides a means to dissect fungal gene expression in infected tissues at single-cell resolution.

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Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC11423565PMC
http://dx.doi.org/10.1128/msphere.00282-24DOI Listing

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