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Determination of Region-Specific Roles of the M3 Muscarinic Acetylcholine Receptor in Gastrointestinal Motility

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Abstract

Background

The specific role of the M3 muscarinic acetylcholine receptor in gastrointestinal motility under physiological conditions is unclear, due to a lack of subtype-selective compounds.

Aims

The objective of this study was to determine the region-specific role of the M3 receptor in gastrointestinal motility.

Methods

We developed a novel positive allosteric modulator (PAM) for the M3 receptor, PAM-369. The effects of PAM-369 on the carbachol-induced contractile response of porcine esophageal smooth muscle and mouse colonic smooth muscle (ex vivo) and on the transit in mouse small intestine and rat colon (in vivo) were examined.

Results

PAM-369 selectively potentiated the M3 receptor under the stimulation of its orthosteric ligands without agonistic or antagonistic activity. Half-maximal effective concentrations of PAM activity for human, mouse, and rat M3 receptors were 0.253, 0.345, and 0.127 μM, respectively. PAM-369 enhanced carbachol-induced contraction in porcine esophageal smooth muscle and mouse colonic smooth muscle without causing any contractile responses by itself. The oral administration of 30 mg/kg PAM-369 increased the small intestinal transit in both normal motility and loperamide-induced intestinal dysmotility mice but had no effects on the colonic transit, although the M3 receptor mRNA expression is higher in the colon than in the small intestine.

Conclusions

This study provided the first direct evidence that the M3 receptor has different region-specific roles in the motility function between the small intestine and colon in physiological and pathophysiological contexts. Selective PAMs designed for targeted subtypes of muscarinic receptors are useful for elucidating the subtype-specific function.

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Abbreviations

AUC:

Area under the curve

[Ca2+]i :

Intracellular concentrations of Ca2+

ChAT:

Choline acetyltransferase

CHO:

Chinese hamster ovary

4-DAMP:

1,1-Dimethyl-4-diphenylacetoxypiperidinium iodide

DMSO:

Dimethyl sulfoxide

EC20 :

20% of maximum effective concentration

EC50 :

Half maximal effective concentration

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

GPCR:

G-protein coupled receptor

5-HT:

5-Hydroxytryptamine

IC50 :

Half maximal inhibitory concentration

ICC:

Intestinal cells of Cajal

KES:

118 MM K + extracellular solution

NES:

137-MM normal extracellular solution

NOS:

Nitric oxide synthase

PAM:

Positive allosteric modulator

VIP:

Vasoactive intestinal polypeptide

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Acknowledgments

The authors thank the laboratory members for their hard work and thorough discussion and Japan Medical Communication (http://www.japan-mc.co.jp) for editing a draft of this manuscript.

Funding

This project has been executed using the institutions’ (Kyushu University and Mochida Pharmaceutical Co., Ltd.) budgets including a grant from Mochida Pharmaceutical Co., Ltd. to EI and YO for this collaborative research. This study was also supported in part by the Japan Society for the Promotion of Science KAKENHI (20K08334 and 22K19530). The funders provided support in the form of salaries for authors YI, CH, and YT (Mochida Pharmaceutical Co., Ltd.).

Author information

Authors and Affiliations

  1. Research Center, Mochida Pharmaceutical Co., Ltd., 722 Uenohara, Jimba, Gotemba, 412-8524, Japan

    Yoko Igarashi-Hisayoshi, Chika Higashi & Yasushi Taguchi

  2. Department of Medicine and Bioregulatory Science, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Yoko Igarashi-Hisayoshi, Eikichi Ihara, Xiaopeng Bai, Hiroko Ikeda, Yoshimasa Tanaka, Haruei Ogino, Takatoshi Chinen & Yoshihiro Ogawa

  3. Department of Gastroenterology and Metabolism, Graduate School of Medical Sciences, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka, 812-8582, Japan

    Eikichi Ihara

  4. Division of Molecular Cardiology, Research Institute of Angiocardiology, Graduate School of Medical Sciences, Kyushu University, Fukuoka, 812-8582, Japan

    Mayumi Hirano

Authors
  1. Yoko Igarashi-Hisayoshi
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Contributions

YI, EI, YT, HO, and TC proposed the research design. YI, XB, CH, and HI conducted experiments. YI and CH performed data analysis. EI, MH, and YT contributed new reagents or analytic tools. YI, EI, XB, YT, HO, and TC wrote or contributed to the writing of the manuscript. YO supervised this study. All authors contributed to the interpretation of the study.

Corresponding author

Correspondence to Yoko Igarashi-Hisayoshi.

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Disclosures

YI, CH, and YT are the employees of Mochida Pharmaceutical Co., Ltd. whose company partly funded this study. The patent of PAM-369 was established and is owned by Mochida Pharmaceutical Co., Ltd.

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Igarashi-Hisayoshi, Y., Ihara, E., Bai, X. et al. Determination of Region-Specific Roles of the M3 Muscarinic Acetylcholine Receptor in Gastrointestinal Motility. Dig Dis Sci 68, 439–450 (2023). https://doi.org/10.1007/s10620-022-07637-y

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