{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,3]],"date-time":"2026-04-03T13:15:50Z","timestamp":1775222150849,"version":"3.50.1"},"reference-count":51,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2020,11,10]],"date-time":"2020-11-10T00:00:00Z","timestamp":1604966400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Neurogastroenterology Motil"],"published-print":{"date-parts":[[2021,4]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>We previously reported the specificity of a novel anti\u2010human peripheral choline acetyltransferase (hpChAT) antiserum for immunostaining of cholinergic neuronal cell bodies and fibers in the human colon. In this study, we investigate 3D architecture of intrinsic cholinergic innervation in the human sigmoid colon and the relationship with nitrergic neurons in the enteric plexus.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We developed a modified CLARITY tissue technique applicable for clearing human sigmoid colon specimens and immunostaining with hpChAT antiserum and co\u2010labeling with neuronal nitric oxide synthase (nNOS) antibody. The Z\u2010stack confocal images were processed for 3D reconstruction\/segmentation\/digital tracing and computational quantitation by Imaris 9.2 and 9.5.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>In the mucosa, a local micro\u2010neuronal network formed of hpChAT\u2010ir fibers and a few neuronal cell bodies were digitally assembled. Three layers of submucosal plexuses were displayed in 3D structure that were interconnected by hpChAT\u2010ir fiber bundles and hpChAT\u2010ir neurons were rarely co\u2010labeled by nNOS. In the myenteric plexus, 30.1% of hpChAT\u2010ir somas including Dogiel type I and II were co\u2010labeled by nNOS and 3 classes of hpChAT\u2010ir nerve fiber strands were visualized in 3D images and videos. The density and intensity values of hpChAT\u2010ir fibers in 3D structure were significantly higher in the circular than in the longitudinal layer.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Inferences<\/jats:title><jats:p>The intrinsic cholinergic innervation in the human sigmoid colon was demonstrated layer by layer for the first time in 3D microstructures. This may open a new venue to assess the structure\u2010function relationships and pathological alterations in colonic diseases.<\/jats:p><\/jats:sec>","DOI":"10.1111\/nmo.14030","type":"journal-article","created":{"date-parts":[[2020,11,11]],"date-time":"2020-11-11T03:56:58Z","timestamp":1605067018000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":13,"title":["Intrinsic cholinergic innervation in the human sigmoid colon revealed using CLARITY, three\u2010dimensional (3D) imaging, and a novel anti\u2010human peripheral choline acetyltransferase (hpChAT) antiserum"],"prefix":"10.1111","volume":"33","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-0067-510X","authenticated-orcid":false,"given":"Pu\u2010Qing","family":"Yuan","sequence":"first","affiliation":[{"name":"UCLA\/Digestive Diseases Research Core Center, Vatche and Tamar Manoukian Digestive Diseases Division, Department of Medicine UCLA David Geffen School of Medicine Los Angeles CA USA"},{"name":"VA Greater Los Angeles Healthcare System Los Angeles CA USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jean\u2010Pierre","family":"Bellier","sequence":"additional","affiliation":[{"name":"Molecular Neuroscience Research Center Shiga University of Medical Science Otsu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Tao","family":"Li","sequence":"additional","affiliation":[{"name":"UCLA\/Digestive Diseases Research Core Center, Vatche and Tamar Manoukian Digestive Diseases Division, Department of Medicine UCLA David Geffen School of Medicine Los Angeles CA USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Mary R.","family":"Kwaan","sequence":"additional","affiliation":[{"name":"Department of Surgery UCLA David Geffen School of Medicine Los Angeles CA USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hiroshi","family":"Kimura","sequence":"additional","affiliation":[{"name":"Molecular Neuroscience Research Center Shiga University of Medical Science Otsu Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yvette","family":"Tach\u00e9","sequence":"additional","affiliation":[{"name":"UCLA\/Digestive Diseases Research Core Center, Vatche and Tamar Manoukian Digestive Diseases Division, Department of Medicine UCLA David Geffen School of Medicine Los Angeles CA USA"},{"name":"VA Greater Los Angeles Healthcare System Los Angeles CA USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2020,11,10]]},"reference":[{"key":"e_1_2_9_2_1","first-page":"7","article-title":"Neuronal populations in the submucous plexus of the human colon","volume":"166","author":"Hoyle CH","year":"1989","journal-title":"J Anat"},{"key":"e_1_2_9_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrgastro.2012.32"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-0897-4_3"},{"key":"e_1_2_9_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-1838(00)00127-2"},{"key":"e_1_2_9_6_1","doi-asserted-by":"publisher","DOI":"10.1117\/12.281502"},{"key":"e_1_2_9_7_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.10.034"},{"key":"e_1_2_9_8_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth.2481"},{"key":"e_1_2_9_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.07.017"},{"key":"e_1_2_9_10_1","doi-asserted-by":"publisher","DOI":"10.1038\/srep34331"},{"key":"e_1_2_9_11_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn.2928"},{"key":"e_1_2_9_12_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2014.03.042"},{"key":"e_1_2_9_13_1","doi-asserted-by":"publisher","DOI":"10.1038\/nmeth1036"},{"key":"e_1_2_9_14_1","doi-asserted-by":"publisher","DOI":"10.1038\/nprot.2012.119"},{"key":"e_1_2_9_15_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.3000231"},{"key":"e_1_2_9_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/0889-1605(86)90004-2"},{"key":"e_1_2_9_17_1","doi-asserted-by":"publisher","DOI":"10.1002\/1097-0185(20010101)262:1<58::AID-AR1011>3.0.CO;2-V"},{"key":"e_1_2_9_18_1","first-page":"67","volume-title":"Physiology of the Gastrointestinal Tract","author":"Wood JD","year":"1987"},{"key":"e_1_2_9_19_1","doi-asserted-by":"publisher","DOI":"10.1172\/JCI102641"},{"key":"e_1_2_9_20_1","unstructured":"StanboroughRJ.Sigmoid Colon: Where It Is What It Does and Why It's Important.https:\/\/www.healthline.com\u203ahealth\u203adigestive\u2010health\u203asigmoid\u2010colon. 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