However, Emc3 depletion has no influence on Hes1 expression examined by qPCR (Supplementary Fig.?3b). endoplasmic reticulum (ER) stress. Mitigating ER stress with tauroursodeoxycholate acid alleviates secretory dysfunction and GW-1100 restores organoid formation. Our study identifies a dominant role of Emc3 in maintaining intestinal mucosal homeostasis. Introduction Intestinal mucosa which consists of physical barrier and luminal immune factors, such as secretory IgA, defensins, lysozyme, Agr2, and Reg3, is the first line of host to defend against external deleterious stimuli.1 The physical barrier is composed of a layer of epithelial cells that classified into absorptive and secretory lineages, both of which are constantly replenished by Lgr5+ intestinal stem cells (ISCs). Absorptive enterocytes are the major type of intestinal epithelial cells, making up to 80% of the entire GW-1100 epithelium. The secretory lineages include mucus-producing goblet cells, antimicrobial peptides (AMPs)-secreting Paneth cells, hormone-secreting enteroendocrine cells, and rare infection-mediating tuft cells.2,3 Both perturbation of mucosal integrity and reduction in innate immune factors are implicated in the pathogenesis of inflammatory bowel diseases (IBDs).4C6 As the most abundant secretory lineage, goblet cells secrete various hydrophilic glycoproteins, including mucins and other protective factors such as Agr2, Zg16, TFF3, FCGBP, and RELM, to form a lubricative barrier blocking microbial invasion into intestinal epithelium.7 These glycoproteins are synthesized in endoplasmic reticulum (ER), followed by selectively packaged into intercellular granules. 8 ER plays dominant functions in ensuring proper folding and maturation of mucins, whose disruption accompanies with goblet cell depletion GW-1100 and spontaneous colitis.9C11 Paneth cells are intermingled with ISCs at the bottom of crypts in the small intestine. Due to its specialized position, Paneth cells constitute a significant component of the ISC niche by supplying requisite signaling ligands (Wnt, EGF, and Dll4), as well as metabolite (Lactate), for ISC maintenance and differentiation.12C14 Paneth cells also serve as a part of the innate immune system by secreting AMPs into gut lumen to influence intestinal microbiome thus to maintain microbiome-host homeostasis.13,15,16 It has been reported that elevated ER stress in gene, is a subunit of the highly GW-1100 conserved ER membrane protein complex (EMC), which is involved FKBP4 in protein folding.23,24 Accumulated evidence shows that EMC ensures the biosynthesis, stabilization and/or trafficking of specific multi-pass membrane proteins in contamination. Deletion of Emc3 in intestinal epithelium impairs mucus-producing function of goblet cells and differentiation of Paneth cells, which modulate gut bacterial composition. Moreover, Emc3 deletion abolishes intestinal organoid culture through disrupting stem cell niche constructed by Paneth cells. Mitigating ER stress caused by Emc3 deficiency with tauroursodeoxycholate acid (TUDCA) rescues secretory lineages and restores the formation of intestinal organoids. Results Emc3 deletion leads to spontaneous inflammation and increased susceptibility to induced colitis We first detected the expression of Emc3 in intestinal mucosa using a specific antibody against N terminal of Emc3 protein (Fig.?1a). We found that Emc3 exhibited a ubiquitous expression pattern along the villus-crypt axis in intestinal epithelium (Fig.?1b), while its expression level was much lower in laminar propria. Open in a separate windows Fig. 1 Emc3 deletion leads to spontaneous inflammation and increased susceptibility to DSS-induced colitis.Pattern of Emc3 expression detected by western blot (a) and immunofluorescence (b) from 8-week-old control and ileum. mice at indicated time points. Data represent mean??SEM. to generate gut epithelium-specific deletion of (mouse pups were GW-1100 born at expected Mendelian frequency and showed no gross phenotype at birth. We validated the efficient depletion of Emc3 at both protein and mRNA level by immunoblots (Fig.?1a) and quantitative PCR (qPCR) (Supplementary Fig.?1b). Immunostaining further verified that Emc3 expression was removed from intestinal epithelium, but not from laminar propria or other tissues (Fig.?1b). Emc3-deficient mice started to display decreased body weight within 2 weeks of age relative to their littermates (Fig.?1c), and such reduction in the growth rate was gradually obvious. mice were used as control, as no defect was observed compared to wild-type mice. and control littermates were used for the following study. There was no.
