Project description:Sequencing of mRNA isolated from the thymus of control mice and mice overexpressing RANKL in keratin 5 (K5)-positive cells for 1 weeks.
Project description:We used embryonic day 15 submandibular salivary glands from K5-Venus mice, which were produced using the bovine-K5 promoter driving Venus expression, N=3 separate experiments
Project description:The circumvallate papillae (CVP) and foliate papillae (FoP) of the posterior tongue contain taste buds in close proximity to specialized salivary glands, known as von Ebner and minor salivary glands, respectively. The developmental relationship between taste buds and these salivary glands remains largely unexplored. Lineage tracing studies in mice have revealed that Lgr5 marks taste bud stem cells. Here, we report single-cell RNA sequencing of the entire CVP and FoP of mice, yielding transcriptional profiles of cells from tongue surface epithelium, taste buds and the associated salivary glands. We unveiled a developmental trajectory in which taste buds, the associated salivary glands and the non-taste tongue surface epithelium originate from a common Lgr5+ cell. We describe long-term organoid culture conditions for these cells and confirm their tripotency at the clonal level in vitro. CVP and FoP harbor chemosensory units consisting of taste bud and salivary gland cells derived from the same parental Lgr5+ stem cell.
Project description:Salivary glands that produce and secret saliva, which is essential for lubrication, digestion, immunity, and oral homeostasis, consist of diverse cells. The long-term maintenance of diverse salivary gland cells in organoids remains problematic. Here, we established long-term murine salivary gland organoids from 3 major salivary glands, including parotid gland (PG), submandibular gland (SMG), and sublingual gland (SLG). Murine salivary gland organoids expressed gland-specific genes and proteins of acinar, myoepithelial, and duct cells. Organoids were maintained in growth media (named GEM) and further underwent differentiation in differentiation media (named DAM). Our study will provide an experimental platform for the exploration of mechanisms involvled in tissue regeneration, development, or several salivary gland diseases.
Project description:Purpose: Next-generation sequencing (NGS) has revolutionized systems-based analysis of cellular functions. The goal of this study is to compare NGS-derived salivary gland transcriptome profilings (RNA-seq) to better understand the molecular nature of the physiological differences in adult murine salivary glands. Methods: Major murine salivary gland mRNA profiles were generated by deep sequencing, in triplicate, using Illumina HiSeq 2000. The sequence reads that passed quality filters were analyzed at the gene level with STAR followed by Cufflinks. In vivo NaCl reabsorption measurements were performed for validation. Results: Using an optimized data analysis workflow, we mapped about 15 million sequence reads per sample to the mouse genome (build mm10) and identified 1991 genes that were differentially expressed across three major salivary glands. RNA-seq data provided valuable insights into the nature of the functional differences among the major salivary glands Conclusions: Our study represents the first detailed analysis of murine salivary gland transcriptomes, with biologic replicates, generated by RNA-seq technology. Our results confirm functions of many genes, identified using genetically modified mice. We conclude that RNA-seq-based transcriptome characterization would offer a comprehensive and sensitive evaluation of the gene expression.
Project description:The circumvallate papillae (CVP) and foliate papillae (FoP) of the posterior tongue contain taste buds in close proximity to specialized salivary glands, known as von Ebner and minor salivary glands, respectively. The developmental relationship between taste buds and these salivary glands remains largely unexplored. Lineage tracing studies in mice have revealed that Lgr5 marks taste bud stem cells. Here, we report single-cell RNA sequencing of the entire CVP and FoP of mice, yielding transcriptional profiles of cells from tongue surface epithelium, taste buds and the associated salivary glands. We unveiled a developmental trajectory in which taste buds, the associated salivary glands and the non-taste tongue surface epithelium originate from a common Lgr5+ cell. We describe long-term organoid culture conditions for these cells and confirm their tripotency at the clonal level in vitro. CVP and FoP harbor chemosensory units consisting of taste bud and salivary gland cells derived from the same parental Lgr5+ stem cell.
Project description:Ticks are notorious carriers of pathogens; these blood-sucking arthropods can spread a variety of deadly diseases. The salivary gland is the main organ in ticks, and this organ begins to develop rapidly when Ixodidae ticks suck blood. When these ticks reach a critical weight, the salivary glands stop developing and begin to degenerate. Specific developmental features of the salivary glands are regulated by multiple factors, such as hormones, proteins and other small molecular substances. In this study, we used iTRAQ quantitative proteomics to study dynamic changes in salivary gland proteins in female Haemaphysalis longicornis at four feeding stages: unfed, partially fed, semi-engorged, and engorged. Through bioinformatics analysis of a large number of proteins, we found that molecular motor- and TCA cycle-related proteins play an important role during the development of the salivary glands. The results of RNAi experiments showed that when dynein, kinesin, isocitrate dehydrogenase, and citrate synthase were knocked down, ticks were unable to suck blood normally. The structure and function of the salivary glands were also significantly affected. In addition, four proteins from H. longicornis were found to have very low homology with those from mammals, including humans. Therefore, it is expected that drugs or antibodies targeting these unique sequences can be designed to kill ticks.
Project description:In this study we have explored the effects of ERdj5 chaperone protein deletion in the proteome of 129SV mouse submandibular salivary glands. This knockout mouse model recapitulates many aspects of Sjögren’s Syndrome, including salivary gland inflammatory infiltrations, anti-nuclear autoantibodies and others (Apostolou et al., Front.Immunol.2019;10,506). Additionally, for 12SV wildtype animals we explore the proteome profile of the extensive sexual dimorphism observed in the submanidular glands of mice.
Project description:Salivary glands that produce and secret saliva, which is essential for lubrication, digestion, immunity, and oral homeostasis, consist of diverse cells. Maintenance of diverse salivary gland cells in organoids remains problematic. Here, we established human salivary gland organoids, which is composed of multiple cellular subsets, from 3 major salivary glands, including parotid gland (PG), submandibular gland (SMG), and sublingual gland (SLG). Human salivary gland organoids expressed gland-specific genes and proteins of acinar, myoepithelial, and duct cells. Organoids were maintained in growth media (named GEM) and further underwent differentiation in differentiation media (named DAM). Our study will provide an experimental platform for the exploration of mechanisms involvled in tissue regeneration, development, or several salivary gland diseases.
Project description:We use mRNA-seq to transcriptionally profile larval salivary gland tissue from Drosophila third instar larvae. These data provide insights into tissue physiology and can be used to identify tissue specific transcripts. Salivary glands were dissected from 200 wandering third instar larvae and the associated fat body was removed.Salivary glands were transferred to Graces unsupplemented medium on ice prior to RNA extraction with TRIzol reagent. mRNA-seq samples were prepared from 10 ug of total RNA and subject to Illumina based sequencing.