{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,7,30]],"date-time":"2025-07-30T12:31:46Z","timestamp":1753878706178,"version":"3.41.2"},"reference-count":29,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2017,11,28]],"date-time":"2017-11-28T00:00:00Z","timestamp":1511827200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100003329","name":"Ministerio de Econom\u00eda y Competitividad","doi-asserted-by":"publisher","award":["IJCI\u20102015\u201026099"],"award-info":[{"award-number":["IJCI\u20102015\u201026099"]}],"id":[{"id":"10.13039\/501100003329","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003042","name":"Else Kr\u00f6ner-Fresenius-Stiftung","doi-asserted-by":"publisher","award":["2013_A29"],"award-info":[{"award-number":["2013_A29"]}],"id":[{"id":"10.13039\/501100003042","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Neurogastroenterology Motil"],"published-print":{"date-parts":[[2018,5]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Alterations of site\u2010specific gene expression profiles in disease\u2010relevant networks within the different layers of the intestinal wall may contribute to the onset and clinical course of gastrointestinal disorders. To date, no systematic analysis has assessed and compared sub\u2010regional gene expression patterns in all distinct layers of the gut using fresh frozen human samples. Our aim was to establish an optimized protocol for site\u2010specific <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> isolation in order to achieve maximum <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> quality and amount for subsequent gene expression analysis combining laser\u2010capture microdissection (<jats:styled-content style=\"fixed-case\">LCM<\/jats:styled-content>) with a probe\u2010based technology, the NanoString <jats:styled-content style=\"fixed-case\">nC<\/jats:styled-content>ounter Analysis system.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Four full\u2010thickness colon samples from patients who underwent surgery due to pathological conditions were processed and separated into epithelium, lamina propria, myenteric plexus, submucosa, and tunica muscularis by <jats:styled-content style=\"fixed-case\">LCM<\/jats:styled-content>. Site\u2010specific marker expression by <jats:styled-content style=\"fixed-case\">nC<\/jats:styled-content>ounter technology was performed on total <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> from each sub\u2010region, respectively.<\/jats:p><\/jats:sec><jats:sec><jats:title>Key Results<\/jats:title><jats:p>Collecting ~10\u00a0mm\u00b2 (~100\u00a0000\u2010250\u00a0000 cells) of tissue from the epithelial layer, lamina propria, and myenteric plexus provided sufficient amounts of <jats:styled-content style=\"fixed-case\">RNA<\/jats:styled-content> of appropriate quality for subsequent analyses. In contrast, ~40\u00a0mm\u00b2 (~250\u00a0000\u2010650\u00a0000 cells) of tissue were dissected from the less cell\u2010rich submucosal and tunica muscularis layer. <jats:styled-content style=\"fixed-case\">nC<\/jats:styled-content>ounter analysis revealed a site\u2010specific expression pattern of marker genes in the different layers of the colonic wall which were highly correlating (<jats:italic>r<\/jats:italic>\u00a0&gt;\u00a0.9).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions and Inferences<\/jats:title><jats:p><jats:styled-content style=\"fixed-case\">LCM<\/jats:styled-content> in combination with <jats:styled-content style=\"fixed-case\">nC<\/jats:styled-content>ounter expression analysis enables site\u2010specific, sensitive, reliable detection, and quantification of <jats:styled-content style=\"fixed-case\">mRNA<\/jats:styled-content> from histologically heterogeneous tissues.<\/jats:p><\/jats:sec>","DOI":"10.1111\/nmo.13261","type":"journal-article","created":{"date-parts":[[2017,11,29]],"date-time":"2017-11-29T17:25:37Z","timestamp":1511976337000},"source":"Crossref","is-referenced-by-count":4,"title":["Site\u2010specific gene expression analysis from archived human intestine samples combining laser\u2010capture microdissection and multiplexed color\u2010coded probes"],"prefix":"10.1111","volume":"30","author":[{"given":"A.","family":"Braun","sequence":"first","affiliation":[{"name":"Department of Human Molecular Genetics Institute of Human Genetics University of Heidelberg  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9368-1356","authenticated-orcid":false,"given":"C.","family":"Martinez","sequence":"additional","affiliation":[{"name":"Department of Human Molecular Genetics Institute of Human Genetics University of Heidelberg  Heidelberg Germany"},{"name":"Digestive System Research Unit Department of Gastroenterology Institut de Recerca Vall d'Hebron Hospital Universitari Vall d'Hebron Universitat Aut\u00f2noma de Barcelona (Departamento de Medicina)  Barcelona Spain"},{"name":"COST Action BM1106 Genes in Irritable Bowel Syndrome Research Network Europe (GENIEUR)  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S.","family":"Schmitteckert","sequence":"additional","affiliation":[{"name":"Department of Human Molecular Genetics Institute of Human Genetics University of Heidelberg  Heidelberg Germany"},{"name":"COST Action BM1106 Genes in Irritable Bowel Syndrome Research Network Europe (GENIEUR)  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"R\u00f6th","sequence":"additional","affiliation":[{"name":"Department of Human Molecular Genetics Institute of Human Genetics University of Heidelberg  Heidelberg Germany"},{"name":"nCounter Core Facility Institute of Human Genetics University of Heidelberg  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3212-1881","authenticated-orcid":false,"given":"F.","family":"Lasitschka","sequence":"additional","affiliation":[{"name":"Institute of Pathology University of Heidelberg  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7881-6112","authenticated-orcid":false,"given":"B.","family":"Niesler","sequence":"additional","affiliation":[{"name":"Department of Human Molecular Genetics Institute of Human Genetics University of Heidelberg  Heidelberg Germany"},{"name":"COST Action BM1106 Genes in Irritable Bowel Syndrome Research Network Europe (GENIEUR)  Heidelberg Germany"},{"name":"nCounter Core Facility Institute of Human Genetics University of Heidelberg  Heidelberg Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2017,11,28]]},"reference":[{"key":"e_1_2_10_2_1","doi-asserted-by":"publisher","DOI":"10.1136\/gutjnl-2016-312518"},{"key":"e_1_2_10_3_1","doi-asserted-by":"publisher","DOI":"10.1038\/ajg.2011.472"},{"key":"e_1_2_10_4_1","doi-asserted-by":"publisher","DOI":"10.1152\/ajpgi.00068.2014"},{"key":"e_1_2_10_5_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0165-0270(03)00232-2"},{"key":"e_1_2_10_6_1","doi-asserted-by":"publisher","DOI":"10.1136\/bmjopen-2015-008439"},{"key":"e_1_2_10_7_1","doi-asserted-by":"publisher","DOI":"10.1007\/s11010-015-2596-7"},{"key":"e_1_2_10_8_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2005.10.010"},{"key":"e_1_2_10_9_1","doi-asserted-by":"publisher","DOI":"10.1007\/s12575-010-9026-8"},{"key":"e_1_2_10_10_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.mam.2005.12.006"},{"key":"e_1_2_10_11_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0053711"},{"key":"e_1_2_10_12_1","doi-asserted-by":"publisher","DOI":"10.1136\/gut.2008.148908"},{"key":"e_1_2_10_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/ibd.21572"},{"key":"e_1_2_10_14_1","doi-asserted-by":"publisher","DOI":"10.1007\/978-1-61779-163-5_15"},{"key":"e_1_2_10_15_1","doi-asserted-by":"publisher","DOI":"10.1586\/14737159.7.5.647"},{"key":"e_1_2_10_16_1","doi-asserted-by":"publisher","DOI":"10.1261\/rna.1261708"},{"key":"e_1_2_10_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ab.2011.05.009"},{"key":"e_1_2_10_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.biochi.2012.06.031"},{"key":"e_1_2_10_19_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jneumeth.2005.04.019"},{"key":"e_1_2_10_20_1","doi-asserted-by":"publisher","DOI":"10.1186\/1471-2164-11-163"},{"key":"e_1_2_10_21_1","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-15-0262"},{"key":"e_1_2_10_22_1","doi-asserted-by":"publisher","DOI":"10.1186\/1472-6750-11-46"},{"key":"e_1_2_10_23_1","doi-asserted-by":"publisher","DOI":"10.1186\/gb-2002-3-7-research0034"},{"key":"e_1_2_10_24_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0005536"},{"key":"e_1_2_10_25_1","doi-asserted-by":"publisher","DOI":"10.1111\/j.1365-2982.2009.01424.x"},{"key":"e_1_2_10_26_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13104-015-1813-5"},{"key":"e_1_2_10_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-013-1690-y"},{"key":"e_1_2_10_28_1","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1385"},{"key":"e_1_2_10_29_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v21.i12.3499"},{"key":"e_1_2_10_30_1","first-page":"88","article-title":"Site\u2010specific gene expression analysis of human intestinal samples by laser capture microdissection combined with nCounter analysis","volume":"27","author":"Niesler B","year":"2015","journal-title":"Neurogastroenterol Motil"}],"container-title":["Neurogastroenterology &amp; Motility"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fnmo.13261","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/nmo.13261","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,23]],"date-time":"2023-09-23T15:03:58Z","timestamp":1695481438000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/nmo.13261"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,11,28]]},"references-count":29,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2018,5]]}},"alternative-id":["10.1111\/nmo.13261"],"URL":"https:\/\/doi.org\/10.1111\/nmo.13261","archive":["Portico"],"relation":{},"ISSN":["1350-1925","1365-2982"],"issn-type":[{"type":"print","value":"1350-1925"},{"type":"electronic","value":"1365-2982"}],"subject":[],"published":{"date-parts":[[2017,11,28]]},"article-number":"e13261"}}