title: Human ENS regulates the intestinal epithelial barrier permeability and a tight junction-associated protein ZO-1 via VIPergic pathways.
abstract: Although the enteric nervous system (ENS) has been shown to regulate various mucosal functions, its role in the physiological control of the human intestinal epithelial barrier is unknown. The aim of this study was to investigate whether the ENS is able to modulate epithelial barrier permeability and a key tight junction-associated protein, zonula occludens-1 (ZO-1). Therefore, we developed a co-culture model, consisting of human submucosa containing the submucosal neuronal network and human polarized colonic epithelial monolayers (HT29-Cl.16E or Caco-2). Submucosal neurons were activated by electrical field stimulation (EFS). Permeability was assessed by measuring the flux of paracellular permeability markers (FITC-dextran or FITC-inulin) across epithelial monolayers. Expression of ZO-1 was determined by immunofluorescence, quantitative immunoblot analysis, and real time RT-PCR. Using the coculture model, we showed that EFS of submucosal neurons resulted in a reduction in FITC-dextran or FITC-inulin fluxes, which was blocked by TTX. In HT29-Cl.16E, the effect of submucosal neuron activation was blocked by a VIP receptor antagonist (VIPra) and reproduced by VIP. Furthermore, ZO-1 expression (mRNA, protein) assessed in HT29-Cl.16E, was significantly increased after submucosal neuron activation by EFS. These effects on ZO-1 expression were blocked by TTX and VIPra and reproduced by VIP. In conclusion, our results strongly suggest a modulatory role of VIPergic submucosal neuronal pathways on intestinal epithelial barrier permeability and ZO-1 expression.
authors.affiliation: Institut National de la Santé et de la Recherche Médicale U 539, Faculté de Médecine, 1, rue Gaston Veil, 44035 Nantes Cedex, France.
authors.forename: Michel
authors.initials: M
authors.lastname: Neunlist
authors.forename: Ferial
authors.initials: F
authors.lastname: Toumi
authors.forename: Tsvetelina
authors.initials: T
authors.lastname: Oreschkova
authors.forename: Marc
authors.initials: M
authors.lastname: Denis
authors.forename: Joel
authors.initials: J
authors.lastname: Leborgne
authors.forename: Christian L
authors.initials: CL
authors.lastname: Laboisse
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Galmiche
authors.forename: Anne
authors.initials: A
authors.lastname: Jarry
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018938:Caco-2 Cells; D002460:Cell Line; D002470:Cell Survival; D018920:Coculture Techniques; D003106:Colon; D046508:Culture Techniques; D004558:Electric Stimulation; D017615:Enteric Nervous System / Q000502:physiology*; D005260:Female; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology / Q000294:innervation* / Q000378:metabolism* / Q000502:physiology; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D008875:Middle Aged; D010539:Permeability; D010750:Phosphoproteins / Q000378:metabolism*; D019108:Tight Junctions / Q000378:metabolism*; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism*; D062826:Zonula Occludens-1 Protein
chemical_list: D008565:Membrane Proteins; D010750:Phosphoproteins; C569042:TJP1 protein, human; D062826:Zonula Occludens-1 Protein; D014660:Vasoactive Intestinal Peptide