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Identification of neurons that express 5-hydroxytryptamine4 receptors in intestine

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Abstract

5-Hydroxytryptamine (5-HT) is an endogenous stimulant of intestinal propulsive reflexes. It exerts its effects partly through 5-HT4 receptors; 5-HT4 receptor agonists that are stimulants of intestinal transit are in clinical use. Both pharmacological and recent immunohistochemical studies indicate that 5-HT4 receptors are present on enteric neurons but the specific neurons that express the receptors have not been determined. In the present work, we describe the characterization of an anti-5-HT4 receptor antiserum that reveals immunoreactivity for enteric neurons and other cell types in the gastrointestinal tract. With this antiserum, 5-HT4 receptor immunoreactivity has been found in the muscularis mucosae of the rat oesophagus, a standard assay tissue for 5-HT4 receptors. It is also present in the muscularis mucosae of the guinea-pig and mouse oesophagus. In guinea-pig small intestine and rat and mouse colon, 5-HT4 receptor immunoreactivity occurs in subpopulations of enteric neurons, including prominent large neurons. Double-staining has shown that these large neurons in the guinea-pig small intestine are also immunoreactive for two markers of intrinsic primary afferent neurons, cytoplasmic NeuN and calbindin. Some muscle motor neurons in the myenteric ganglia are immunoreactive for this receptor, whereas it is rarely expressed by secretomotor neurons. Immunoreactivity also occurs in the interstitial cells of Cajal but is faint in the external muscle. Expression of the protein and mRNA has been confirmed in extracts containing enteric neurons. The observations suggest that one site of action of 5-HT4 receptor agonists is the intrinsic primary afferent neurons.

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Acknowledgement

We thank Dr. Philip Thompson of the Department of Pharmaceutical Biology and Pharmacology, Monash University, for the synthesis of the peptide used to raise the antibodies.

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Authors and Affiliations

  1. Department of Anatomy & Cell Biology and Centre for Neuroscience, University of Melbourne, Parkville 3010, Australia

    Daniel P. Poole, Bo Xu, Shir Lin Koh, Billie Hunne & John B. Furness

  2. Department of Pharmaceutical Biology and Pharmacology, Monash University, Parkville 3052, Australia

    Ian M. Coupar & Helen R. Irving

  3. Discovery Biology Research Group, Pfizer Global Research and Development, Nagoya Laboratories, Pfizer, Aichi, Japan

    Katsuhiro Shinjo

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  1. Daniel P. Poole
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  2. Bo Xu
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  3. Shir Lin Koh
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Correspondence to John B. Furness.

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This work was supported by the National Health and Medical Research Council of Australia and Pfizer Pharmaceuticals, Japan.

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Poole, D.P., Xu, B., Koh, S.L. et al. Identification of neurons that express 5-hydroxytryptamine4 receptors in intestine. Cell Tissue Res 325, 413–422 (2006). https://doi.org/10.1007/s00441-006-0181-9

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  • DOI: https://doi.org/10.1007/s00441-006-0181-9

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