Department of Microbiology and Molecular Genetics, McGovern Medical School, The University of Texas Health Science Center at Houston, Houston, Texas, USA.
mSphere. 2024 Sep 25;9(9):e0028224. doi: 10.1128/msphere.00282-24. Epub 2024 Aug 22.
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.
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.
从感染器官中获得的微生物基因表达测量对于理解发病机制非常有价值。然而，目前的方法仅限于“总体”分析，忽略了微生物细胞的异质性和病变的空间结构。在这里，我们报告了使用杂交链反应 RNA 荧光杂交（HCR RNA-FISH）在感染小鼠的舌头上以单细胞分辨率可视化和定量转录物。该方法与固定冷冻和福尔马林固定石蜡包埋组织兼容。我们记录了编码与口腔念珠菌病相关产物的 mRNA 的细胞间变异和有趣的时空表达模式。该方法为宿主相互作用的基因表达分析提供了空间维度。
是一种真菌共生体，栖息在人体的多个粘膜表面。免疫抑制、抗生素诱导的微生物失调或植入的医疗设备会损害粘膜完整性，使过度生长和传播，导致粘膜疾病，如口咽念珠菌病或危及生命的全身感染。分析在感染的粘膜或任何其他感染的器官中表达的真菌基因对于理解发病机制至关重要。理想情况下，这些转录谱测量应该以单细胞水平揭示任何基因的表达。然而，目前方法通常达到的分辨率将大多数基因表达测量限制在细胞群体平均值上。本报告中描述的方法提供了一种在单细胞分辨率下剖析感染组织中真菌基因表达的手段。
