Project description:The enteric nervous system (ENS) and SIP (Smooth muscle cells-Interstitial cells of Cajal-PDGFRα+ cells) syncytium play an important role in controlling gastrointestinal motility. This study aimed to investigate the dynamic regulatory mechanisms of the ENS-SIP system on colon motility during postnatal development. Colonic samples of postnatal 1 week (PW1), PW3, and PW5 old murine were characterized by RNA sequencing, qRT-PCR, and immunoblotting. The current study showed that ENS and glial cells including ICCs and PDGFRα+ cells become increasingly mature in both the postnatal proximal and distal colon. The transcriptional expression of Pdgfrα, c-Kit, P2ry1, Nos1, and Slc18a3, and the protein expression of nNOS, c-Kit, and ANO1 significantly increased with age. In conclusion, during postnatal development, molecular data demonstrated the gradual maturation of ICC and PDGFRα+ cells, including nitrergic and cholinergic nerves and purinergic receptors. Our findings are important in understanding the role of ENS and interstitial cells with specific receptors for the generation of mature colonic migrating motor complexes in young murine colons.
Project description:Genome scale expression data on absolute numbers of gene isoforms offer essential clues for cellular functions and biological processes. Gastrointestinal (GI) motility is regulated by smooth muscle cells (SMC) closely contacted with interstitial cells of Cajal (ICC) and fibroblast-like PDGFRα+ cells (PαC), forming an electrical syncytium. To uncover genetic identifies and cellular functions of the cells, we isolated these three cell populations from mouse small intestine and colon, obtained the transcriptome for each type of cells, and built each cell type transcriptome browser. To our knowledge, this is the first genetic resource providing a comprehensive reference for all mRNA transcripts expressed in these unique GI cell populations. Integration of these data with the UCSC genome browser revealed novel cell-specific markers, ion channel and transporter isoforms, and unique cellular and biological functions of these cells in GI physiology. Our transcriptome browsers bring new insight into the alternative expression of genes in different types of the cells and provides references for future functional studies. mRNA profiles of SMC, ICC and PDGFRa cells isolated from mouse jejunum and colon were generated by deep sequencing using Illumina Hiseq2000..
Project description:Genome scale expression data on absolute numbers of gene isoforms offer essential clues for cellular functions and biological processes. Gastrointestinal (GI) motility is regulated by smooth muscle cells (SMC) closely contacted with interstitial cells of Cajal (ICC) and fibroblast-like PDGFRα+ cells (PαC), forming an electrical syncytium. To uncover genetic identifies and cellular functions of the cells, we isolated these three cell populations from mouse small intestine and colon, obtained the transcriptome for each type of cells, and built each cell type transcriptome browser. To our knowledge, this is the first genetic resource providing a comprehensive reference for all mRNA transcripts expressed in these unique GI cell populations. Integration of these data with the UCSC genome browser revealed novel cell-specific markers, ion channel and transporter isoforms, and unique cellular and biological functions of these cells in GI physiology. Our transcriptome browsers bring new insight into the alternative expression of genes in different types of the cells and provides references for future functional studies.
Project description:To map gene regulation downstream of cholesterol overload and NF-kappaB signaling in smooth muscle cells (SMCs), we cultured primary aortic SMCs from wildtype mice with cyclodextrin-complexed cholesterol or the prototypical NF-kappaB activator, tumor necrosis factor (TNF), or both.
Project description:We report the application of single-cell based sequencing for high throughput profiling of the mouse urinary bladder. By utilizing multiple dissociation techniques and library preparation techniques we generated an atlas comprising 43,119 cells including major cell types absent from previous reports. We found that previous single-cell profiling of the mouse bladder lacked a major cell type in the detrusor smooth muscle and incorrectly annotated other cell types such as mesothelial cells. Using the atlas, we elucidated aspects of bladder biology including urothelial differentiation, the identity of interstitial cells of Cajal, detrusor smooth muscle control and immune distributions. Finally, we combine the single-cell based sequencing with spatial transcriptomics and imaging mass cytometry to add spatial context to transcriptomic profiling.
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from Mus musculus tissues (Heart, Liver, Lung, Kidney, Skeletal Muscle, Thymus)
Project description:SILAC based protein correlation profiling using size exclusion of protein complexes derived from seven Mus musculus tissues (Heart, Brain, Liver, Lung, Kidney, Skeletal Muscle, Thymus)