Non‐neuronal, but atropine‐sensitive ileal contractile responses to short‐chain fatty acids: age‐dependent desensitization and restoration under inflammatory conditions in mice

Intestinal epithelial cells sense short‐chain fatty acids (SCFAs) to secrete non‐neuronal acetylcholine (ACh). However, the roles of luminal SCFAs and epithelial ACh under normal and pathological conditions remain unknown. We examined ileal contractile responses to SCFAs at different ages and their mucosal cholinergic alterations under inflammatory conditions. Ileal contractile responses to SCFAs in 1‐day‐old pups to 7‐week‐old mice were compared using an isotonic transducer, and responses to an intraperitoneal injection of lipopolysaccharide (LPS) were analyzed in 7‐week‐old mice. The mRNA expression levels of a SCFA activate free fatty acid receptor, acetylcholinesterase (AChE), choline acetyltransferase (Chat), and choline transporter‐like protein 4 (CTL4) were measured using real‐time quantitative RT‐PCR. AChE was analyzed by histochemical and optical enzymatic assays. Atropine‐sensitive ileal contractile responses to SCFAs occurred in all 1‐day‐old pups, but were frequently desensitized after the weaning period. These contractile responses were not inhibited by tetrodotoxin and did not appear when the mucosal layer had been scraped off. Contractile desensitization in 7‐week‐old mice was abolished in the presence of the AChE inhibitor, eserine, which was consistent with increased AChE activity after weaning. Ileal contractions to SCFAs in adult mice were restored by LPS, which significantly increased the epithelial mRNA expression of Chat and CTL4. Atropine‐sensitive ileal contractile responses to SCFAs constitutively occur in the newborn period, and are desensitized during developmental stages following the up‐regulated expression of AChE in the villous mucosa, but are restored under inflammatory conditions possibly via the release of epithelial ACh.