{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,16]],"date-time":"2026-06-16T11:50:22Z","timestamp":1781610622045,"version":"3.54.5"},"reference-count":71,"publisher":"Wiley","issue":"10","license":[{"start":{"date-parts":[[2014,9,3]],"date-time":"2014-09-03T00:00:00Z","timestamp":1409702400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK093525"],"award-info":[{"award-number":["DK093525"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Neurogastroenterology Motil"],"published-print":{"date-parts":[[2014,10]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Anterograde <jats:italic>in vitro<\/jats:italic> tracing of the pelvic nerve (<jats:styled-content style=\"fixed-case\">PN<\/jats:styled-content>) and visualization in the horizontal plane in whole mount preparations has been fundamental in the analysis of distribution of peripheral nerves innervating the colorectum. Here, we performed a similar analysis, but in cryostat sections of the mouse colorectum, allowing for a more direct visualization of nerve distribution in all tissue layers.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Colorectum with attached <jats:styled-content style=\"fixed-case\">PN<\/jats:styled-content>s was dissected from adult male BalbC mice. Presence of active afferents was certified by single fiber recording of fine <jats:styled-content style=\"fixed-case\">PN<\/jats:styled-content> fibers. This was followed by \u2018bulk\u2019 (all fibers) anterograde tracing using biotinamide (<jats:styled-content style=\"fixed-case\">BTA<\/jats:styled-content>). Histo\u2010 and immunohistochemical techniques were used for visualization of <jats:styled-content style=\"fixed-case\">BTA<\/jats:styled-content>\u2010positive nerves, and evaluation of co\u2010localization with calcitonin gene\u2010related peptide (<jats:styled-content style=\"fixed-case\">CGRP<\/jats:styled-content>), respectively. Tissue was analyzed using confocal microscopy on transverse or longitudinal colorectum sections.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>Abundant <jats:styled-content style=\"fixed-case\">BTA<\/jats:styled-content>\u2010positive nerves spanning all layers of the mouse colorectum and contacting myenteric plexus neurons, distributing within the muscle layer, penetrating deeper into the organ and contacting blood vessels, submucosal plexus neurons or even penetrating the mucosa, were regularly detected. Several traced axons co\u2010localized <jats:styled-content style=\"fixed-case\">CGRP<\/jats:styled-content>, supporting their afferent nature. Finally, anterograde tracing of the <jats:styled-content style=\"fixed-case\">PN<\/jats:styled-content> also exposed abundant <jats:styled-content style=\"fixed-case\">BTA<\/jats:styled-content>\u2010positive nerves in the major pelvic ganglion.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions &amp; Inferences<\/jats:title><jats:p>We present the patterns of innervation of extrinsic axons across layers in the mouse colorectum, including the labile mucosal layer. The proposed approach could also be useful in the analysis of associations between morphology and physiology of peripheral nerves targeting the different layers of the colorectum.<\/jats:p><\/jats:sec>","DOI":"10.1111\/nmo.12419","type":"journal-article","created":{"date-parts":[[2014,9,3]],"date-time":"2014-09-03T07:20:03Z","timestamp":1409728803000},"page":"1494-1507","source":"Crossref","is-referenced-by-count":6,"title":["Distribution across tissue layers of extrinsic nerves innervating the mouse colorectum \u2013 An <i>in vitro<\/i> anterograde tracing study"],"prefix":"10.1111","volume":"26","author":[{"given":"Pablo R.","family":"Brumovsky","sequence":"first","affiliation":[{"name":"School of Biomedical Sciences Austral University Pilar Buenos Aires Argentina"},{"name":"CONICET (Consejo Nacional de Investigaciones Cient\u00edficas y T\u00e9cnicas) Buenos Aires Argentina"},{"name":"Pittsburgh Center for Pain Research Department of Anesthesiology University of Pittsburgh 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