{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,8]],"date-time":"2026-07-08T06:11:31Z","timestamp":1783491091623,"version":"3.55.0"},"reference-count":32,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T00:00:00Z","timestamp":1698278400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100014498","name":"Chemistry, Engineering and Medicine for Human Health, Stanford University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100014498","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100004410","name":"European Molecular Biology Organization","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100004410","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100018128","name":"Ludwig Family Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100018128","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100010319","name":"Shurl and Kay Curci Foundation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100010319","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100014373","name":"Wu Tsai Neurosciences Institute, Stanford University","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100014373","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Neurogastroenterology Motil"],"published-print":{"date-parts":[[2024,1]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:sec>\n                    <jats:title>Background<\/jats:title>\n                    <jats:p>Accurately reporting the identity and representation of enteric nervous system (ENS) neuronal subtypes along the length of the gastrointestinal (GI) tract is critical to advancing our understanding of ENS control of GI function. Reports of varying proportions of subtype marker expression have employed different dissection techniques to achieve wholemount muscularis preparations of myenteric plexus. In this study, we asked whether differences in GI dissection methods could introduce variability into the quantification of marker expression.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Methods<\/jats:title>\n                    <jats:p>We compared three commonly used methods of ENS wholemount dissection: two flat\u2010sheet preparations that differed in the order of microdissection and fixation and a third rod\u2010mounted peeling technique. We also tested a reversed orientation variation of flat\u2010sheet peeling, two step\u2010by\u2010step variations of the rod peeling technique, and whole\u2010gut fixation as a tube. We assessed marker expression using immunohistochemistry, genetic reporter lines, confocal microscopy, and automated image analysis.<\/jats:p>\n                  <\/jats:sec>\n                  <jats:sec>\n                    <jats:title>Key Results and Conclusions<\/jats:title>\n                    <jats:p>We found no significant differences between the two flat\u2010sheet preparation methods in the expression of calretinin or neuronal nitric oxide synthase (nNOS) as a proportion of total neurons in ileum myenteric plexus. However, the rod\u2010mounted peeling method resulted in decreased proportion of neurons labeled for both calretinin and nNOS. This method also resulted in decreased transgenic reporter fluorescent protein (tdTomato) for substance P in distal colon and choline acetyltransferase (ChAT) in both ileum and distal colon. These results suggest that labeling among some markers, both native protein and transgenic fluorescent reporters, is decreased by the rod\u2010mounted mechanical method of peeling. The step\u2010by\u2010step variations of this method point to mechanical manipulation of the tissue as the likely cause of decreased labeling. Our study thereby demonstrates a critical variability in wholemount muscularis dissection methods.<\/jats:p>\n                  <\/jats:sec>","DOI":"10.1111\/nmo.14693","type":"journal-article","created":{"date-parts":[[2023,10,26]],"date-time":"2023-10-26T05:50:45Z","timestamp":1698299445000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Comparison of wholemount dissection methods for neuronal subtype marker expression in the mouse myenteric plexus"],"prefix":"10.1111","volume":"36","author":[{"given":"Julieta","family":"Gomez\u2010Frittelli","sequence":"first","affiliation":[{"name":"Department of Chemical Engineering Stanford University  Stanford California USA"},{"name":"Wu Tsai Neurosciences Institute Stanford University  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryan","family":"Hamnett","sequence":"additional","affiliation":[{"name":"Wu Tsai Neurosciences Institute Stanford University  Stanford California USA"},{"name":"Department of Neurosurgery Stanford University School of Medicine  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia A.","family":"Kaltschmidt","sequence":"additional","affiliation":[{"name":"Wu Tsai Neurosciences Institute Stanford University  Stanford California USA"},{"name":"Department of Neurosurgery Stanford University School of Medicine  Stanford California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2023,10,26]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrgastro.2012.32"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.7554\/eLife.42914"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-008-0684-7"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41593-020-00736-x"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2020.08.003"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-004-0895-5"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1002\/cne.10948"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2018.06.021"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1053\/j.gastro.2020.09.032"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0306-4522(83)90001-5"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-021-82785-1"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.18632\/aging.101677"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2022.08.030"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00493164"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2011.01.010"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncir.2014.00076"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1038\/nn.2467"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-020-03543-6"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1038\/nrgastro.2016.107"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41575-020-0271-2"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0171239"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1111\/nmo.12114"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-4939-7244-9_19"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jss.2013.02.044"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2022.799717"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-022-03608-x"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2019.12.002"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.abd6176"},{"key":"e_1_2_10_30_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1619406114"},{"key":"e_1_2_10_31_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2020.109012"},{"key":"e_1_2_10_32_1","doi-asserted-by":"publisher","DOI":"10.1007\/BF00318629"},{"key":"e_1_2_10_33_1","doi-asserted-by":"publisher","DOI":"10.3390\/biomedicines9050465"}],"container-title":["Neurogastroenterology &amp; 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