Pharmacological characterization of purinergic inhibitory neuromuscular transmission in the human colon

Authors

  • D. Gallego,

    1. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, (CIBERehd), Instituto de Salud Carlos III, Barcelona, Spain
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    • Both these authors have contributed equally to this study.

  • V. Gil,

    1. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
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    • Both these authors have contributed equally to this study.

  • J. Aleu,

    1. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
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  • M. Martinez-Cutillas,

    1. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
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  • P. Clavé,

    1. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, (CIBERehd), Instituto de Salud Carlos III, Barcelona, Spain
    2. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
    3. Fundació de Gastroenterologia Dr Vilardell and Department of Surgery, Hospital de Mataró, Mataró, Barcelona, Spain
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  • M. Jimenez

    1. Centro de Investigación Biomédica en Red de Enfermedades Hepáticas y Digestivas, (CIBERehd), Instituto de Salud Carlos III, Barcelona, Spain
    2. Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Barcelona, Spain
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Address for Correspondence
Marcel Jiménez VMD, PhD, Department of Cell Biology, Physiology and Immunology, Edifici V, Universitat Autònoma de Barcelona, 08193 Bellaterra, Spain.
Tel: +34 93 581 1566; fax: +34 93 581 2006;
e-mail: marcel.jimenez@uab.es

Abstract

Background  In the present study, we further characterize the purinergic receptors mediating the inhibitory junction potential (IJP) and smooth muscle relaxation in the human colon using a new, potent and selective agonist (MRS2365), and antagonists (MR2279 and MRS2500) of the P2Y1 receptor. The P2Y12 antagonist AR-C66096 was tested as well. Using this pharmacological approach, we tested whether β-nicotinamide adenine dinucleotide (β-NAD) fulfilled the criteria to be considered an inhibitory neurotransmitter in the human colon.

Methods  We carried out muscle bath and microelectrode experiments on circular strips from the human colon and calcium imaging recordings on HEK293 cells, which constitutively express the human P2Y1 receptor.

Key Results  Both the fast component of IJP and non-nitrergic relaxation was concentration-dependently inhibited by MRS2279 and MRS2500. This antagonism was confirmed in HEK293 cells. However, AR-C66096 did not modify either inhibitory response. Adenosine 5′-Ο-2-thiodiphosphate and MRS2365 caused a smooth muscle hyperpolarization and transient inhibition of spontaneous motility that was antagonized by MRS2279 and MRS2500. β-Nicotinamide adenine dinucleotide inhibited the spontaneous motility (IC50 = 3.3 mmol L−1). Nevertheless, this effect was not antagonized by high concentrations of P2Y1 antagonists.

Conclusions & Inferences  Inhibitory purinergic neuromuscular transmission in the human colon was pharmacologically assessed by the use of new P2Y1 receptor antagonists MRS2179, MRS2279, and MRS2500. The rank order of potency of the P2Y1 antagonists is MRS2500 > MRS2279 > MRS2179. We found that β-NAD partially fulfills the criteria to be considered an inhibitory neurotransmitter in the human colon, but the relative contribution of each purine (ATP/ADP vsβ-NAD) requires further studies.

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