Project description:As the largest salivary gland in oral cavity, the parotid gland plays an important role in initial digesting and lubricating food. The abnormal secretory function of parotid gland can lead to dental caries and oral mucosal inflammation. In recent years, single-cell RNA sequencing (scRNA-seq) has been used to explore the heterogeneity and diversity of cells in various organs and tissues. However, the transcription profile of human parotid gland at single-cell resolution has not been reported yet. In this study, we constructed the cell atlas of human parotid gland using 10x Genomics platform. Characteristic gene analysis identified the biological functions of serous acinar cell populations in secreting digestive enzymes and antibacterial proteins. We revealed the specificity and similarity of parotid gland comparing to other digestive glands through comparative analyses of other published scRNA-seq datasets. We also identified the cell-specific expression of hub genes for Sjogren’s syndrome in human parotid gland by integrating the results of GWAS and bulk RNA-seq, which highlighted the importance of immune cell dysfunction in parotid Sjogren’s syndrome pathogenesis.
Project description:Human synovial Single cell RNA-seq was performed on three tissue samples from healthy donors. This experiment was done to explore the heterogeneity of cells in healthy human synovial joint and enabled the comparison of cellular states and composition to those of publicly available single cell RNA-seq datasets from psoriatic arthritis and rheumatoid arthritis patients. Human synovial cells were loaded immediately after tissue dissociation with up to 25,000 cells in a single well of a Chromium chip G (10x Genomics). 3’ gene expression libraries were generated using Chromium Next GEM Single Cell 3' Kit 3.1 with 3' Feature Barcode Kit and dual indexing (10x Genomics protocol CG000316 Rev C). Libraries were sequenced as paired end (PE) 150 bp by Illumina sequencing to 65-80% saturation. Reads were mapped to the GRCh38 human genome (GENCODE) using the 10x Genomics Cell Ranger pipeline (7.2.0).
Project description:Although mesenchyme is essential for inducing the epithelium of ectodermal organs, its precise role in organ-specific epithelial fate determination remains poorly understood. To elucidate roles of tissue interactions in cellular differentiation, we performed single cell RNA sequencing and imaging analyses of recombined tissues in which embryonic mouse salivary gland mesenchyme and epithelium were switched ex vivo. We found partial induction of molecules that define gland-specific acinar and myoepithelial cells in recombined salivary epithelium. Parotid epithelium (serous gland) recombined with submandibular mesenchyme (mixed serous-mucous, but predominantly mucous gland) began to express mucous acinar genes not intrinsic to the parotid gland. While myoepithelial cells do not normally line parotid acini, newly induced myoepithelial cells densely populated recombined parotid acini. However, mucous acinar and myoepithelial markers continued to be expressed in submandibular epithelial cells recombined with parotid mesenchyme. Consequently, some epithelial cells appeared to be plastic, such that their fate could still be altered in response to mesenchymal signaling, whereas other epithelial cells appeared to be already committed to a specific fate. We also discovered evidence for bidirectional induction: transcriptional changes were observed not only in the epithelium but also in the mesenchyme after heterotypic tissue recombination. For example, parotid epithelium induced the expression of muscle-related genes in submandibular fibroblasts that began to mimic parotid fibroblast gene expression. These studies provide the first comprehensive unbiased molecular characterization of tissue recombination approaches exploring the regulation of cell fate.
Project description:Although mesenchyme is essential for inducing the epithelium of ectodermal organs, its precise role in organ-specific epithelial fate determination remains poorly understood. To elucidate roles of tissue interactions in cellular differentiation, we performed single cell RNA sequencing and imaging analyses of recombined tissues in which embryonic mouse salivary gland mesenchyme and epithelium were switched ex vivo. We found partial induction of molecules that define gland-specific acinar and myoepithelial cells in recombined salivary epithelium. Parotid epithelium (serous gland) recombined with submandibular mesenchyme (mixed serous-mucous, but predominantly mucous gland) began to express mucous acinar genes not intrinsic to the parotid gland. While myoepithelial cells do not normally line parotid acini, newly induced myoepithelial cells densely populated recombined parotid acini. However, mucous acinar and myoepithelial markers continued to be expressed in submandibular epithelial cells recombined with parotid mesenchyme. Consequently, some epithelial cells appeared to be plastic, such that their fate could still be altered in response to mesenchymal signaling, whereas other epithelial cells appeared to be already committed to a specific fate. We also discovered evidence for bidirectional induction: transcriptional changes were observed not only in the epithelium but also in the mesenchyme after heterotypic tissue recombination. For example, parotid epithelium induced the expression of muscle-related genes in submandibular fibroblasts that began to mimic parotid fibroblast gene expression. These studies provide the first comprehensive unbiased molecular characterization of tissue recombination approaches exploring the regulation of cell fate.
Project description:We applied both single cell and bulk RNA-sequencing to characterize and identify uniqueness in gene expression of the mouse Parotid gland compared to other tissues including the submandibular gland
Project description:Gene expression of 4 human parotid gland samples were comapred with 4 mouse parotid gland samples. Human and mouse parotid gland gene expression was screened using custom-designed targeted cDNA containing probes for 198 genes which encode for ion/water transporter (75 genes) and receptor/regulatory (101 genes) proteins potentially involved in the fluid secretion mechanism, as well as 11 secretory protein genes and 11 control genes