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Transient receptor potential vanilloid 4 inhibits mouse colonic motility by activating NO-dependent enteric neurotransmission

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Abstract

Recent studies implicate TRPV4 receptors in visceral pain signaling and intestinal inflammation. Our aim was to evaluate the role of TRPV4 in the control of gastrointestinal (GI) motility and to establish the underlying mechanisms. We used immunohistochemistry and PCR to study TRPV4 expression in the GI tract. The effect of TRPV4 activation on GI motility was characterized using in vitro and in vivo motility assays. Calcium and nitric oxide (NO) imaging were performed to study the intracellular signaling pathways. Finally, TRPV4 expression was examined in the colon of healthy human subjects. We demonstrated that TRPV4 can be found on myenteric neurons of the colon and is co-localized with NO synthase (NOS-1). In vitro, the TRPV4 agonist GSK1016790A reduced colonic contractility and increased inhibitory neurotransmission. In vivo, TRPV4 activation slowed GI motility and reduced stool production in mouse models mimicking pathophysiological conditions. We also showed that TRPV4 activation inhibited GI motility by reducing NO-dependent Ca2+ release from enteric neurons. In conclusion, TRPV4 is involved in the regulation of GI motility in health and disease.

Key messages

• Recent studies implicate TRPV4 in pain signaling and intestinal inflammation.

• Our aim was to characterize the role of TRPV4 in the control of GI motility.

• We found that TRPV4 activation reduced colonic contractility.

• Our studies also showed altered TRPV4 mRNA expression in IBS-C patients.

• TRPV4 may be a novel pharmacological target in functional GI diseases.

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Abbreviations

1400W:

N-(3-Aminomethyl) benzylacetamidine, NOS-2 blocker

EFS:

Electrical field stimulation

GFAP:

Glial fibrillary acidic protein

GSK1016790A:

(N-((1S)-1-{[4-((2S)-2-{[(2,4-Dichlorophenyl)sulfonyl]amino}-3-hydroxypropanoyl)-1-piperazinyl]carbonyl}-3-methylbutyl)-1-benzothiophene-2-carboxamide, TRPV4 agonist

IBS:

Irritable bowel syndrome

LMMP:

Longitudinal muscle–myenteric plexus

NANC:

Non-adrenergic, non-cholinergic

ODQ:

1H-[1,2,4]Oxadiazolo[4,3-a]quinoxalin-1-one, ​soluble guanylate cyclase inhibitor

PTIO:

2-Phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide, NO scavenger

RN 1734:

2,4-Dichloro-N-isopropyl-N-(2-isopropylaminoethyl)benzenesulfonamide, TRPV4 antagonist

RuR:

Ruthenium red, TRPV antagonist

SB 366791:

4′-Chloro-3-methoxycinnamanilide, selective TRPV1 antagonist

sGC:

Soluble guanylate cyclase

SMTC:

S-Methyl-l-thiocitrulline, NOS-1 blocker

TRPV4:

Transient receptor potential vanilloid type 4

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Acknowledgments

The authors wish to thank Dr. Kevin Thorneloe (GlaxoSmithKline Pharmaceuticals, King of Prussia, PA, USA) for providing GSK1016790A for this study, Wolfgang Liedtke (Duke University, Durham, NC, USA) for providing the TRPV4−/− mice, and Drs. Catherine Legraverend (Institut de Génomique Fonctionnelle, Montpellier, France) and Richard Dyck (University of Calgary, AB, Canada) for supplying the S100β-GFP mice. SJM was supported by the Dr. T. Chen Fong Doctoral Scholarship in Neuroscience through the Hotchkiss Brain Institute and a Studentship from Alberta Innovates-Health Solutions (AI-HS). KAS is the Crohn’s and Colitis Foundation of Canada Chair in IBD Research. Supported by the Iuventus Plus program of the Polish Ministry of Science and Higher Education (0107/IP1/2013/72 to JF), Medical University of Lodz (503/1-156-04/503-01 to JF), National Science Centre (UMO-2013/11/B/NZ7/01301 and UMO-2014/13/B/NZ4/01179 to JF), NHMRC (454858 to DPP and 63303 to NWB), NIH (DK57840 to NWB), Monash University (to NWB), the Canadian Institutes of Health Research (to KAS), the Deutsche Forschungsgemeinschaft (STO 645/6-1 to MS), and the University of Calgary Research Grant Committee (to MS).

Author contributions

Concept and design: JF, DPP, SJM, KAS, MAS

Analysis and interpretation of data: JF, DPP, NV, SJM, DS, AM, PKZ, AIC, JPT

Drafting the manuscript: JF, SJM, KAS, MAS

Final approval of the version to be published: all authors

Disclosures

The authors have nothing to disclose.

Author information

Authors and Affiliations

  1. Snyder Institute for Chronic Diseases, University of Calgary, Calgary, AB, Canada

    J. Fichna, K. A. Sharkey & M. A. Storr

  2. Department of Medicine, Division of Gastroenterology, University of Calgary, Calgary, AB, Canada

    J. Fichna & M. A. Storr

  3. Department of Physiology & Pharmacology, University of Calgary, Calgary, AB, Canada

    S. J. MacEachern & K. A. Sharkey

  4. Hotchkiss Brain Institute, University of Calgary, Calgary, AB, Canada

    S. J. MacEachern & K. A. Sharkey

  5. Department of Biochemistry, Medical University of Lodz, Lodz, Poland

    J. Fichna

  6. Department of Digestive Tract Diseases, Faculty of Medicine, Medical University of Lodz, Lodz, Poland

    A. Mokrowiecka & E. Małecka-Panas

  7. Monash Institute of Pharmaceutical Sciences, Parkville, Victoria, Australia

    D. P. Poole, N. Veldhuis & N. W. Bunnett

  8. Department of Anatomy and Neuroscience, The University of Melbourne, Parkville, VIC, Australia

    D. P. Poole

  9. Department of Pharmacology, The University of Melbourne, Parkville, VIC, Australia

    N. W. Bunnett

  10. II Medizinische Klinik, Technische Universität München, Munich, Germany

    D Saur

  11. Department of Cytobiochemistry, Faculty of Biology and Environmental Protection, University of Lodz, Lodz, Poland

    P. K. Zakrzewski, A. I. Cygankiewicz & W. M. Krajewska

  12. Center for Translational Neuroscience, Duke University, Durham, NC, USA

    W. Liedtke

  13. Department of Dermatology and Surgery, University of California San Francisco, San Francisco, CA, USA

    M. S. Steinhoff

  14. Laboratory of Cell Biology and Histology, Department of Veterinary Sciences, University of Antwerp, Antwerp, Belgium

    J-P. Timmermans

  15. Department of Medicine, Division of Gastroenterology, Ludwig Maximilians University of Munich, Marchioninistrasse 15, 81377, Munich, Germany

    M. A. Storr

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  1. J. Fichna
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  2. D. P. Poole
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  3. N. Veldhuis
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  4. S. J. MacEachern
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  5. D Saur
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  10. W. M. Krajewska
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Corresponding author

Correspondence to M. A. Storr.

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Fichna, J., Poole, D.P., Veldhuis, N. et al. Transient receptor potential vanilloid 4 inhibits mouse colonic motility by activating NO-dependent enteric neurotransmission. J Mol Med 93, 1297–1309 (2015). https://doi.org/10.1007/s00109-015-1336-5

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  • DOI: https://doi.org/10.1007/s00109-015-1336-5

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