Project description:This study was undertaken to identify gene expression changes in enteric neurons of Hirschsprung disease mice. Methods: We performed single-cell RNA sequencing of enteric nervous system cells isolated from the small intestine of Ednrb-knockout mice at age P14. Results: Through comparisons with published datasets of WT small intestine enteric neurons, we identify a missing neuronal population in the ganglionated small intestine of Hirschsprung disease mice. Conclusion: Single-cell RNA sequencing permits identification of cellular perturbations in Hirschsprung disease models.
Project description:Identify cytotoxic CD4+ T cells in human intestine and analyze characteristics of the population in IBD patients by using single cell RNA-seq
Project description:We here systematically studied the interaction network of small intestine crypt cells.We first created a reference of cell types based on single-cell mRNA sequencing, which we used to create virtual doublets and triplets. These virtual combinations were then used to train a machine learning algorithm (random forests) in order to infer the cell types present in the actual picked doublets or triplets. We then counted the number of interactions in between defferent cell types, and compared it to label permution simulation to identify interactions that happens more or less than expected by chance.
Project description:The mammalian intestine is a self-renewing organ that processes ingested food, absorbing nutrients from a sea of biochemical, mechanical, and pathogenic insults. To understand the developmental specification of diverse cell types in the small intestine, we performed time-resolved single-cell transcriptomics across five stages intestinal development: embryonic days (E) 12.5, 14.5, 16.5, and 18.5, and postnatal day (P) 1.5. By dissociating cells from whole organ preparations, we captured the full range of cellular diversity across all intestinal cell lineages. This allowed us to reconstruct the mesenchymal differentiation trajectory and identify the specific genetic programs that give rise to the diverse musculature of the gut wall and villus stroma.
Project description:The study aimed to make a molecular definition of myenteric neuron classes in the mouse small intestine and describe their differentiation from enteric stem cells during embryogenesis. Method: We performed single cell RNA-sequencing of enteric neurons from small intestine at the juvenile stage (postnatal day (P)21) and ENS cells from the small intestine at embryonic day (E)15.5 and E18.5. We confirmed novel marker genes for all classes using immunohistochemistry and characterised the morphology/axons of three of newly discovered ENS neuron types. Results: 12 unique enteric neuron classes (ENC1-12) were identified and confirmed in native tissue by histochemical methods. The communication machinery (ligand, receptors, adhesion molecules), axonal patterns and morphologies were presented to infer functional roles of the ENCs. Analysis of embryonic transcriptomes revealed that enteric stem cells differentiate into two neuronal branches, that successively diversify into ENC1-7 and ENC8-12, respectively. Conclusion: Utilizing single cell RNA-sequencing we conclude that murine small intestine consists of 12 major myenteric classes that emerge through a step-wise fashion during embryonic development. Reference: Morarach, Mikhailova et al., 2020. BioRxiv 2020.03.02.955757
Project description:Small intestine is highly innervated by nociceptor neurons. Single cell RNA sequencing (scRNA-seq) of ileal epithelial cells was performed to analyze the role of nociceptor neurons in regulating small intestinal epithelial cells.