DOI: 10.5061/dryad.jwstqjqgr
title: Colonic cytokines and chemokines in DNBS colitis

description: Abdominal pain in irritable bowel syndrome and inflammatory bowel disease
 is thought to be driven by processes that sensitize sensory nerves
 innervating the gut. How sensory nerves become sensitized is not clear,
 but their terminals in the gut are surrounded by enteric glia. Here, we
 tested the hypothesis that intercellular enteric glia-to-nociceptor
 signaling contributes to visceral hypersensitivity during inflammation. In
 vivo and in vitro models of acute inflammation were used in combination
 with protein and RNA labeling, and cellular assays of activity and
 mediator release. Mechanisms of interaction between glia and nociceptors
 were studied using Trpv1Cre;GCaMP5gtdT;GFAP-hM3Dq mice, in which glial
 activity is controlled by chemogenetics while simultaneously recording
 nociceptor activity using calcium imaging. Mice lacking glial connexin-43
 were used in combination with visceromotor reflex recordings to disrupt
 glial intercellular signaling and study its impact on visceral
 sensitivity. Acute colitis induces a transient increase in proinflammatory
 cytokines including IL-1β, which is produced in part by glia and
 facilitates glial connexin-43 function. Provoking glial activity under
 these conditions changes the normal benign influence of glia on
 nociceptors to one where glia have a sensitizing effect on gut-innervating
 nociceptors. The mechanisms responsible for glial-driven visceral
 hypersensitivity involve an upregulation of glial COX-2 and an increase in
 stimulated glial PGE2 release which acts on nociceptor EP4 receptors. In
 vivo recordings show that colonic IL-1β shifts normal innocuous stimuli
 toward a noxious range through mechanisms that require glial connexin-43.
 Enteric glia sensitize gut nociceptors during inflammation. Cell-specific
 therapies targeting the glial mechanisms identified here could benefit
 treatments for visceral pain.
description: Whole colon samples were collected from control and DNBS treated
 mice at 6 hours, 48 hours, and 3 weeks. Tissue was processed for multiplex
 immunoassay as previously described (<em>50</em>). Colonic
 samples were homogenized in a Tris-buffered saline Tween buffer solution
 to extract protein. After, homogenates were centrifuged at
 10,000<em>g</em> for 10 minutes at 4°C. Cytokine/chemokine
 levels were evaluated using bead-based multiplex immunoassays (Eve
 Technologies, Calgary, AB, Canada).
description: # Colonic cytokines and chemokines in DNBS colitis
 [https://doi.org/10.5061/dryad.jwstqjqgr](https://doi.org/10.5061/dryad.jwstqjqgr) Analysis of colonic cytokine and chemokine levels during the the time course of DNBS colitis in male and female mice. Concentrations reflect levels in samples of whole colon in healthy controls, during the acute, proinflammatory phase of colitis (6 hours and 48 hours) and following resolution (3 weeks). Colon homogenates were analyzed Eve Technologies (Calgary, AB) using a commercial bead-based multiplex immunoassay. ## Description of the data and file structure The file contains a spreadsheet of raw data. Individual cytokines are labeled in columns. Samples identified under "Treatment" represent whole colon homogenates obtained from animals at 6 hours post DNBS (6h DNBS), 48 hours post DNBS (48h DNBS), or 3 weeks post DNBS (3 3 DNBS). Animal sex is indicated by "F" for female and "M" for male. "FI" signifies the Fluorescence Intensity of the bead population for a specific analyte. The "Obs Conc" is the observed concentration that corresponds to the FI calculated with the standard curve. OOR is when the FI was outside the range of the standard curve and can't be extrapolated. Cells labeled "ND" are where FI was not detectable. ## Sharing/Access information Data was derived from the following sources: * Morales-Soto et al. Science Signaling