Abstract
Acetylcholine (ACh) is a neurotransmitter that is present in central, parasympathetic, and neuromuscular synapses of mammals. However, non-neuronal ACh is also predicted to function as a local cell signaling molecule. The physiological significance of the presence of non-neuronal ACh in the intestine remains unclear. Here, experiments using cultured crypt–villus organoids that lack nerve and immune cells led us to suggest that endogenous ACh is synthesized in the intestinal epithelium to evoke growth and differentiation of the organoids through activation of muscarinic ACh receptors (mAChRs). Extracts of cultured organoids exhibited a noticeable capacity for ACh synthesis that was sensitive to a potent inhibitor of choline acetyltransferase. Treatment of organoids with carbachol downregulated growth of organoids and expression of marker genes for each epithelial cell type. On the other hand, mAChR antagonists enhanced growth and differentiation of Lgr5-positive stem cells. Collectively, our data provide evidence that endogenous ACh released from mouse intestinal epithelium maintains the homeostasis of intestinal epithelial cell growth and differentiation via mAChRs.
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References
Sastry BV, Sadavongvivad C (1978) Cholinergic systems in non-nervous tissues. Pharmacol Rev 30:65–132
Horiuchi Y, Kimura R, Kato N, Fujii T, Seki M, Endo T et al (2003) Evolutional study on acetylcholine expression. Life Sci 72:1745–1756
Wessler I, Kilbinger H, Bittinger F, Kirkpatrick CJ (2001) The biological role of non-neuronal acetylcholine in plants and humans. Jpn J Pharmacol 85:2–10
Takahashi T, Hamaue N (2010) Molecular characterization of Hydra acetylcholinesterase and its catalytic activity. FEBS Lett 584:511–516
Wessler I, Kirkpatrick CJ (2008) Acetylcholine beyond neurons: the non-neuronal cholinergic system in humans. Br J Pharmacol 154:1558–1571
Cheng H, Leblond CP (1974) Origin, differentiation and renewal of the four main epithelial cell types in the mouse small intestine. V. Unitarian theory of the origin of the four epithelial cell types. Am J Anat 141:537–561
Barker N, van Es JH, Kuipers J, Kujala P, van den Born M, Cozijnsen M et al (2007) Identification of stem cells in small intestine and colon by marker gene Lgr5. Nature 449:1003–1007
Sato T, Vries RG, Snippert HJ, van de Wetering M, Barker N, Stange DE et al (2009) Single Lgr5 stem cells build crypt-villus structures in vitro without a mesenchymal niche. Nature 459:262–265
Takahashi T, Ohnishi H, Sugiura Y, Honda K, Suematsu M, Kawasaki T et al (2014) Non-neuronal acetylcholine as an endogenous regulator of proliferation and differentiation of Lgr5-positive stem cells in mice. FEBS J 281:4672–4690
Acknowledgement
This work was supported by a Grant-in-Aid for Scientific Research (C) (Grant number 26440184) from the Japan Society for the Promotion of Science (JSPS).
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Takahashi, T. (2016). New Trends and Perspectives in the Function of Non-neuronal Acetylcholine in Crypt–Villus Organoids in Mice. In: Turksen, K. (eds) Organoids. Methods in Molecular Biology, vol 1576. Humana, New York, NY. https://doi.org/10.1007/7651_2016_1
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DOI: https://doi.org/10.1007/7651_2016_1
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-7616-4
Online ISBN: 978-1-4939-7617-1
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