title: P2Y Receptors Sensitize Mouse and Human Colonic Nociceptors.
abstract: Activation of visceral nociceptors by inflammatory mediators contributes to visceral hypersensitivity and abdominal pain associated with many gastrointestinal disorders. Purine and pyrimidine nucleotides (e.g., ATP and UTP) are strongly implicated in this process following their release from epithelial cells during mechanical stimulation of the gut, and from immune cells during inflammation. Actions of ATP are mediated through both ionotropic P2X receptors and metabotropic P2Y receptors. P2X receptor activation causes excitation of visceral afferents; however, the impact of P2Y receptor activation on visceral afferents innervating the gut is unclear. Here we investigate the effects of stimulating P2Y receptors in isolated mouse colonic sensory neurons, and visceral nociceptor fibers in mouse and human nerve-gut preparations. Additionally, we investigate the role of Nav1.9 in mediating murine responses. The application of UTP (P2Y2 and P2Y4 agonist) sensitized colonic sensory neurons by increasing action potential firing to current injection and depolarizing the membrane potential. The application of ADP (P2Y1, P2Y12, and P2Y13 agonist) also increased action potential firing, an effect blocked by the selective P2Y1 receptor antagonist MRS2500. UTP or ADP stimulated afferents, including mouse and human visceral nociceptors, in nerve-gut preparations. P2Y1 and P2Y2 transcripts were detected in 80% and 56% of retrogradely labeled colonic neurons, respectively. Nav1.9 transcripts colocalized in 86% of P2Y1-positive and 100% of P2Y2-positive colonic neurons, consistent with reduced afferent fiber responses to UTP and ADP in Na(v)1.9(-/-) mice. These data demonstrate that P2Y receptor activation stimulates mouse and human visceral nociceptors, highlighting P2Y-dependent mechanisms in the generation of visceral pain during gastrointestinal disease.
authors.affiliation: Wingate Institute of Neurogastroenterology, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AJ, United Kingdom, National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: James R F
authors.initials: JR
authors.lastname: Hockley
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: Michael M
authors.initials: MM
authors.lastname: Tranter
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: Cian
authors.initials: C
authors.lastname: McGuire
authors.affiliation: Wingate Institute of Neurogastroenterology, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AJ, United Kingdom, National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: George
authors.identifier: http://orcid.org/0000-0003-1020-5029
authors.initials: G
authors.lastname: Boundouki
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: Vincent
authors.initials: V
authors.lastname: Cibert-Goton
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: Mohamed A
authors.initials: MA
authors.lastname: Thaha
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: L Ashley
authors.initials: LA
authors.lastname: Blackshaw
authors.affiliation: Centre for Neuroscience and Trauma, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AT, United Kingdom, and.
authors.forename: Gregory J
authors.identifier: http://orcid.org/0000-0002-5035-3918
authors.initials: GJ
authors.lastname: Michael
authors.affiliation: Centre for Neuroscience and Trauma, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AT, United Kingdom, and.
authors.forename: Mark D
authors.initials: MD
authors.lastname: Baker
authors.affiliation: National Centre for Bowel Research and Surgical Innovation, and.
authors.forename: Charles H
authors.initials: CH
authors.lastname: Knowles
authors.affiliation: Neusentis (Pfizer Ltd.), Cambridge CB21 6GS, United Kingdom.
authors.forename: Wendy J
authors.initials: WJ
authors.lastname: Winchester
authors.affiliation: Wingate Institute of Neurogastroenterology, Blizard Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London E1 2AJ, United Kingdom, National Centre for Bowel Research and Surgical Innovation, and d.bulmer@qmul.ac.uk.
authors.forename: David C
authors.identifier: http://orcid.org/0000-0002-4703-7877
authors.initials: DC
authors.lastname: Bulmer
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002478:Cells, Cultured; D003106:Colon / Q000187:drug effects / Q000378:metabolism*; D005260:Female; D005727:Ganglia, Spinal / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008875:Middle Aged; D062558:NAV1.9 Voltage-Gated Sodium Channel / Q000502:physiology; D009619:Nociceptors / Q000378:metabolism*; D011685:Purine Nucleotides / Q000494:pharmacology; D011742:Pyrimidine Nucleotides / Q000494:pharmacology; D058470:Receptors, Purinergic P2Y / Q000096:biosynthesis*; D013045:Species Specificity
chemical_list: D062558:NAV1.9 Voltage-Gated Sodium Channel; D011685:Purine Nucleotides; D011742:Pyrimidine Nucleotides; D058470:Receptors, Purinergic P2Y; C491885:Scn11a protein, mouse
keywords: ATP; Nav1.9; P2Y receptors; nociceptor; purinergic signaling; visceral pain