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This study aimed to identify where the sensory information relayed by splanchnic and pelvic afferents integrates within the brainstem. Microinjection of transneuronal viral tracer (herpes simplex virus\u20101 H129 strain expressing EGFP, H129\u2010EGFP) into the distal colon was used to assess the brainstem structures receiving ascending input from the colorectum. H129\u2010EGFP+ cells were distributed in structures involved in ascending sensory relay, descending pain modulation, and autonomic regulation in the medulla from 96\u2009h and in the pontine and caudal midbrain at 120\u2009h after inoculation. In a separate cohort of mice, in\u00a0vivo noxious colorectal distension (CRD) followed by brainstem immunolabeling for phosphorylated MAP kinase ERK 1\/2 (pERK) determined neurons activated by CRD. Many of the structures containing H129\u2010EGFP+ labeling also contained pERK\u2010labeled neurons, indicating H129\u2010EGFP+ labeling in colorectal signaling pathways. Surgical removal of dorsal root ganglia (DRG) containing the cell bodies of splanchnic colorectal afferent neurons significantly reduced CRD\u2010evoked pERK neuronal activation within the caudal ventrolateral medulla, rostral ventromedial medulla, and the lateral parabrachial nuclei. Surgical removal of the DRG containing the cell bodies of pelvic colorectal afferent neurons significantly reduced CRD\u2010evoked pERK neuronal activation within the rostral ventromedial medulla, lateral parabrachial nuclei, the locus coeruleus, Barrington's nucleus, and periaqueductal gray. Collectively, this study showed that the two spinal afferent pathways innervating the colorectum relay information into different brainstem structures and provide new insight into their unique roles in relaying information into the gut\u2010brain axis controlling colorectal sensory\u2010motor function.<\/jats:p>\n                  <jats:p>\n                    <jats:boxed-text content-type=\"graphic\" position=\"anchor\">\n                      <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc70211-gra-0001-m.png\">\n                        <jats:alt-text>image<\/jats:alt-text>\n                      <\/jats:graphic>\n                    <\/jats:boxed-text>\n                  <\/jats:p>","DOI":"10.1111\/jnc.70211","type":"journal-article","created":{"date-parts":[[2025,9,2]],"date-time":"2025-09-02T02:49:51Z","timestamp":1756781391000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Splanchnic and Pelvic Spinal Afferent Pathways Relay Sensory Information From the Mouse Colorectum Into Distinct Brainstem Circuits"],"prefix":"10.1111","volume":"169","author":[{"given":"QingQing","family":"Wang","sequence":"first","affiliation":[{"name":"Visceral Pain Research Group, Hopwood Centre for Neurobiology, Lifelong Health Theme, South Australian Health and Medical Research Institute (SAHMRI)  Adelaide South Australia Australia"},{"name":"College of Medicine and Public Health, Flinders Health and Medical Research Institute Flinders University  Adelaide South Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alice 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