title: Epithelial expression and function of trypsin-3 in irritable bowel syndrome.
abstract: Proteases are key mediators of pain and altered enteric neuronal signalling, although the types and sources of these important intestinal mediators are unknown. We hypothesised that intestinal epithelium is a major source of trypsin-like activity in patients with IBS and this activity signals to primary afferent and enteric nerves and induces visceral hypersensitivity.
  
  
  
  Trypsin-like activity was determined in tissues from patients with IBS and in supernatants of Caco-2 cells stimulated or not. These supernatants were also applied to cultures of primary afferents. mRNA isoforms of trypsin (PRSS1,2and3) were detected by reverse transcription-PCR, and trypsin-3 protein expression was studied by western blot analysis and immunohistochemistry. Electrophysiological recordings and Ca2+imaging in response to trypsin-3 were performed in mouse primary afferent and in human submucosal neurons, respectively. Visceromotor response to colorectal distension was recorded in mice administered intracolonically with trypsin-3.
  
  
  
  We showed that stimulated intestinal epithelial cells released trypsin-like activity specifically from the basolateral side. This activity was able to activate sensory neurons. In colons of patients with IBS, increased trypsin-like activity was associated with the epithelium. We identified that trypsin-3 was the only form of trypsin upregulated in stimulated intestinal epithelial cells and in tissues from patients with IBS. Trypsin-3 was able to signal to human submucosal enteric neurons and mouse sensory neurons, and to induce visceral hypersensitivity in vivo, all by a protease-activated receptor-2-dependent mechanism.
  
  
  
  In IBS, the intestinal epithelium produces and releases the active protease trypsin-3, which is able to signal to enteric neurons and to induce visceral hypersensitivity.
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Claire
authors.initials: C
authors.lastname: Rolland-Fourcade
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Alexandre
authors.initials: A
authors.lastname: Denadai-Souza
authors.affiliation: Laboratory for Enteric Neuroscience (LENS), TARGID, University of Leuven, Leuven, Belgium.
authors.forename: Carla
authors.initials: C
authors.lastname: Cirillo
authors.affiliation: Gastrointestinal Diseases Research Unit, , General Hospital, Queen's University School of Medicine, Kingston, Ontario, Canada.
authors.forename: Cintya
authors.initials: C
authors.lastname: Lopez
authors.affiliation: Gastrointestinal Diseases Research Unit, , General Hospital, Queen's University School of Medicine, Kingston, Ontario, Canada.
authors.forename: Josue Obed
authors.initials: JO
authors.lastname: Jaramillo
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Cleo
authors.initials: C
authors.lastname: Desormeaux
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Cenac
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Motta
authors.affiliation: Oppenheimer Family Center for Neurobiology of Stress and Resilience and CURE: Digestive Diseases Research Center, Vatche and Tamar Manoukian Division of Digestive Diseases, UCLA David Geffen School of Medicine, Los Angeles, California, USA.
authors.forename: Muriel
authors.initials: M
authors.lastname: Larauche
authors.affiliation: Oppenheimer Family Center for Neurobiology of Stress and Resilience and CURE: Digestive Diseases Research Center, Vatche and Tamar Manoukian Division of Digestive Diseases, UCLA David Geffen School of Medicine, Los Angeles, California, USA.
authors.forename: Yvette
authors.initials: Y
authors.lastname: Taché
authors.affiliation: Laboratory for Enteric Neuroscience (LENS), TARGID, University of Leuven, Leuven, Belgium.
authors.forename: Pieter
authors.initials: P
authors.lastname: Vanden Berghe
authors.affiliation: Inserm, UMR913, Nantes, France.|Nantes University, Nantes, France.|Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Neunlist
authors.affiliation: Inserm, UMR913, Nantes, France.|Nantes University, Nantes, France.|Institut des Maladies de l'Appareil Digestif, IMAD, CHU Nantes, Hopital Hôtel-Dieu, Nantes, France.
authors.forename: Emmanuel
authors.initials: E
authors.lastname: Coron
authors.affiliation: Department of Internal Medicine and Digestive Diseases, Pole Digestif, CHU Toulouse, Toulouse, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Kirzin
authors.affiliation: Department of Internal Medicine and Digestive Diseases, Pole Digestif, CHU Toulouse, Toulouse, France.
authors.forename: Guillaume
authors.initials: G
authors.lastname: Portier
authors.affiliation: Department of Internal Medicine and Digestive Diseases, Pole Digestif, CHU Toulouse, Toulouse, France.
authors.forename: Delphine
authors.initials: D
authors.lastname: Bonnet
authors.affiliation: Department of Internal Medicine and Digestive Diseases, Pole Digestif, CHU Toulouse, Toulouse, France.
authors.forename: Laurent
authors.initials: L
authors.lastname: Alric
authors.affiliation: Gastrointestinal Diseases Research Unit, , General Hospital, Queen's University School of Medicine, Kingston, Ontario, Canada.
authors.forename: Stephen
authors.initials: S
authors.lastname: Vanner
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.
authors.forename: Celine
authors.initials: C
authors.lastname: Deraison
authors.affiliation: IRSD, Université de Toulouse, INSERM, INRA, ENVT, UPS, Toulouse, France.|Department of Physiology and Pharmacology, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Vergnolle
mesh_terms: D000818:Animals; D018938:Caco-2 Cells; D016022:Case-Control Studies; D003106:Colon / Q000201:enzymology / Q000294:innervation; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004151:Dipeptides / Q000494:pharmacology; D017615:Enteric Nervous System / Q000166:cytology / Q000000981:diagnostic imaging / Q000187:drug effects; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology*; D005260:Female; D005727:Ganglia, Spinal / Q000166:cytology; D006801:Humans; D006967:Hypersensitivity / Q000201:enzymology; D007413:Intestinal Mucosa / Q000187:drug effects / Q000201:enzymology* / Q000473:pathology; D043183:Irritable Bowel Syndrome / Q000201:enzymology* / Q000235:genetics*; D007555:Isoxazoles / Q000494:pharmacology; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D051379:Mice; D018613:Microscopy, Confocal; D009475:Neurons, Afferent / Q000187:drug effects / Q000502:physiology; D010539:Permeability / Q000187:drug effects; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D044464:Receptor, PAR-2 / Q000037:antagonists & inhibitors / Q000378:metabolism; D014357:Trypsin / Q000235:genetics* / Q000378:metabolism* / Q000494:pharmacology; D014362:Trypsinogen / Q000235:genetics; D015854:Up-Regulation
chemical_list: C553268:5-isoxazoyl-cyclohexylalanyl-isoleucyl-spiroindane-1,4'-piperidine; D017077:Culture Media, Conditioned; D004151:Dipeptides; D007555:Isoxazoles; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D044464:Receptor, PAR-2; C064831:PRSS2 protein, human; D014362:Trypsinogen; C489303:PRSS1 protein, human; C040749:PRSS3 protein, human; C000622735:PRSS3 protein, mouse; D014357:Trypsin
keywords: ABDOMINAL PAIN; IRRITABLE BOWEL SYNDROME; NERVE - GUT INTERACTIONS; TRYPSIN