{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T14:45:30Z","timestamp":1785422730439,"version":"3.56.0"},"posted":{"date-parts":[[2026,2,2]]},"group-title":"Neuroscience","reference-count":45,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T00:00:00Z","timestamp":1769990400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"award":["R01DE029951"],"award-info":[{"award-number":["R01DE029951"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"award":["RM1DE033491"],"award-info":[{"award-number":["RM1DE033491"]}],"id":[{"id":"https:\/\/ror.org\/001d55x84","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000005","name":"United States Department of Defense","doi-asserted-by":"crossref","award":["W81XWH2210239"],"award-info":[{"award-number":["W81XWH2210239"]}],"id":[{"id":"10.13039\/100000005","id-type":"DOI","asserted-by":"crossref"}]},{"name":"CIHR, Canada"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,2]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Proteases and histamine, co-secreted by mast cells and bacteria, sensitize colonic nociceptors and contribute to irritable bowel syndrome (IBS) pain.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Objective<\/jats:title>\n                  <jats:p>\n                    To determine whether irreversible proteolytic cleavage of protease-activated receptor-2 (PAR\n                    <jats:sub>2<\/jats:sub>\n                    ) and its continued activity in endosomes amplify and sustain otherwise transient pronociceptive actions of histamine receptors (HRs) to cause recurrent pain, the defining symptom of IBS.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Design<\/jats:title>\n                  <jats:p>\n                    We investigated the coexpression of PAR\n                    <jats:sub>2<\/jats:sub>\n                    and H\n                    <jats:sub>1<\/jats:sub>\n                    R in nociceptors using RNAscope\n                    <jats:italic>in situ<\/jats:italic>\n                    hybridization and assessed the consequences of coactivation using electrophysiological assays of nociceptor sensitization and biophysical measurements of receptor and effector activity.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    PAR\n                    <jats:sub>2<\/jats:sub>\n                    and H\n                    <jats:sub>1<\/jats:sub>\n                    R were coexpressed by human and mouse dorsal root ganglion nociceptors. Intracolonic infusion of fecal supernatants from IBS patients enhanced mechanosensitivity of colonic nociceptors in mice. Antagonists of PAR\n                    <jats:sub>2<\/jats:sub>\n                    or H\n                    <jats:sub>1-4<\/jats:sub>\n                    R abolished this response. Combined administration of subthreshold concentrations of trypsin and histamine replicated the effects of fecal supernatant and caused hyperexcitability of isolated nociceptors. Pre-activation of PAR\n                    <jats:sub>2<\/jats:sub>\n                    sensitized histamine-induced hyperexcitability. Endocytosis inhibitors prevented this hypersensitivity, consistent with sustained endosomal signaling of PAR\n                    <jats:sub>2<\/jats:sub>\n                    and persistent nociceptor hyperexcitability. Trypsin amplified histamine-induced activation of H\n                    <jats:sub>1<\/jats:sub>\n                    R and \u03b2-arrestin2 and G\u03b1q effectors at the plasmalemma and in endosomes. Conversely, histamine did not sensitize trypsin-induced hyperexcitability of neurons, in line with the inability of histamine to induce sustained nociceptor hypersensitivity.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>\n                    By amplifying and maintaining the otherwise transient actions of H\n                    <jats:sub>1<\/jats:sub>\n                    R and possibly other pain receptors, persistent PAR\n                    <jats:sub>2<\/jats:sub>\n                    endosomal signaling makes a dominant contribution to IBS-related colonic pain.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Summary box<\/jats:title>\n                  <jats:sec>\n                    <jats:title>What is already known on this topic<\/jats:title>\n                    <jats:p>Proteases and histamine are increased in IBS patients and cause visceral pain.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>What this study adds<\/jats:title>\n                    <jats:p>\n                      Prolonged intracellular PAR\n                      <jats:sub>2<\/jats:sub>\n                      signaling sensitizes and maintains H\n                      <jats:sub>1<\/jats:sub>\n                      R activity to amplify and maintain pain.\n                    <\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>How this might affect research, practice or policy<\/jats:title>\n                    <jats:p>\n                      Although neuroactive factors can act synergistically to amplify and maintain IBS pain, antagonists of dominant receptors (\n                      <jats:italic>e<\/jats:italic>\n                      .\n                      <jats:italic>g<\/jats:italic>\n                      ., PAR\n                      <jats:sub>2<\/jats:sub>\n                      ) can provide effective treatment.\n                    <\/jats:p>\n                  <\/jats:sec>\n                <\/jats:sec>","DOI":"10.64898\/2026.01.30.702691","type":"posted-content","created":{"date-parts":[[2026,2,2]],"date-time":"2026-02-02T22:05:57Z","timestamp":1770069957000},"source":"Crossref","is-referenced-by-count":0,"title":["ENDOSOMAL SIGNALING OF PROTEASE-ACTIVATED RECEPTOR-2 AMPLIFIES HISTAMINE-INDUCED PAIN OF IRRITABLE BOWEL SYNDROME"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6595-1884","authenticated-orcid":false,"given":"Nestor N","family":"Jimenez-Vargas","sequence":"first","affiliation":[{"name":"Department of Molecular Pathobiology and the Pain Research Center, College of Dentistry, New York University, New York, New York, 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Pain Research Center, College of Dentistry, New York University, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Brian L.","family":"Schmidt","sequence":"additional","affiliation":[{"name":"Department of Molecular Pathobiology and the Pain Research Center, College of Dentistry, New York University, New York, New York, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David E","family":"Reed","sequence":"additional","affiliation":[{"name":"Gastrointestinal Diseases Research Unit, Kingston General Hospital, Queen\u2019s University, Kingston, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan E","family":"Lomax","sequence":"additional","affiliation":[{"name":"Gastrointestinal Diseases Research Unit, Kingston General Hospital, Queen\u2019s University, Kingston, Ontario, Canada"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen 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