Muscarinic receptor M3 contributes to intestinal stem cell maintenance via EphB/ephrin-B signaling

Life Sci Alliance. 2021 Jul 9;4(9):e202000962. doi: 10.26508/lsa.202000962. Print 2021 Sep.


Acetylcholine (ACh) signaling through activation of nicotinic and muscarinic ACh receptors regulates expression of specific genes that mediate and sustain proliferation, differentiation, and homeostasis in the intestinal crypts. This signaling plays a pivotal role in the regulation of intestinal stem cell function, but the details have not been clarified. Here, we performed experiments using type 3 muscarinic acetylcholine receptor (M3) knockout mice and their intestinal organoids and report that endogenous ACh affects the size of the intestinal stem niche via M3 signaling. RNA sequencing of crypts identified up-regulation of the EphB/ephrin-B signaling pathway. Furthermore, using an MEK inhibitor (U0126), we found that mitogen-activated protein kinase/extracellular signal-regulated kinase (MAPK/ERK) signaling, which is downstream of EphB/ephrin-B signaling, is activated in M3-deficient crypts. Collectively, M3, EphB/ephrin-B, and the MAPK/ERK signaling cascade work together to maintain the homeostasis of intestinal epithelial cell growth and differentiation following modifications of the cholinergic intestinal niche.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biomarkers
  • Cell Differentiation / genetics
  • Cell Proliferation
  • Cell Self Renewal / genetics*
  • Female
  • Fluorescent Antibody Technique
  • Gene Expression
  • Immunohistochemistry
  • Intestinal Mucosa / cytology
  • Intestinal Mucosa / metabolism
  • Intestines / cytology*
  • MAP Kinase Signaling System
  • Male
  • Mice
  • Mice, Knockout
  • Models, Biological
  • Organoids
  • Receptor, Muscarinic M3 / genetics*
  • Receptor, Muscarinic M3 / metabolism*
  • Receptors, Eph Family / metabolism*
  • Signal Transduction*
  • Stem Cells / cytology*
  • Stem Cells / metabolism*


  • Biomarkers
  • Receptor, Muscarinic M3
  • Receptors, Eph Family