Project description:We have employed single cell RNA-sequencing to create a cellular atlas of the female mouse urethra. The epithelial lining of the urethra is an understudied mucosal barrier. Emerging evidence points to the role of the urethra in pathogen surveillance, sensation and response. To investigate the heterogeneity of the resident immune niche in the adult female mouse urethra, we performed single cell RNA-sequencing which revealed the presence of distinct epithelial-associated and stromal macrophages in the urethra.
Project description:We have employed single cell RNA-sequencing to create a cellular atlas of the female mouse urethra. The epithelial lining of the urethra is an understudied mucosal barrier. Emerging evidence points to the role of the urethra in pathogen surveillance, sensation and response. In this study, we instilled female mice transurethrally with PBS or LPS (1 mg/ml) and collected urethras at 3 hours post-instillation. Single cell RNA-sequencing was performing to assess changes in immune and epithelial cell types after LPS treatment.
Project description:The epithelial lining of the urethra is an understudied mucosal barrier with emerging roles in antimicrobial defense during urinary tract infections. Here, we present a comprehensive cellular atlas of the adult female mouse urethra, focusing on epithelial and resident immune cells which act in concert at mucosal sites to direct host defense. Single cell RNA-sequencing revealed two distinct macrophage populations compartmentalized within the epithelium and stroma. Epithelial-associated macrophages display a highly dendritic morphology and populate the urethral lining in increasing numbers over the course of development. Epithelial-associated macrophages, express Cx3cr1, MHCII genes, Cd74 and Aif1/Iba-1, representing an activated macrophage type (Mac-Activated) and also express genes involved in antigen presentation and inflammatory response. In contrast, stromal macrophages express the scavenging receptors Mrc1/Cd206, Lyve1, Cd163 and Mgl2/Cd301b and display a signature enriched for endocytic function, vasculature development and tissue homeostasis (Mac-Endocytic). We identified epithelial cells in the urethral lining and associated glands expressing the monocyte chemoattractant genes Cx3cl1 and Cxcl17. Chemoattractant gene expression in the urethral epithelium follows a proximal-distal gradient which correlates with the increasing density of epithelial-associated macrophages expressing the receptor Cx3cr1, along the proximal-distal axis of the urethra. The study delineates spatially compartmentalized macrophage subsets in the urethra and implicate epithelial-derived chemokines in the establishment of macrophage positioning and functional specialization along the urethral axis.