{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,27]],"date-time":"2026-08-27T06:59:09Z","timestamp":1787813949220,"version":"build-2784847793"},"reference-count":47,"publisher":"The Company of Biologists","issue":"11","license":[{"start":{"date-parts":[[2025,6,3]],"date-time":"2025-06-03T00:00:00Z","timestamp":1748908800000},"content-version":"vor","delay-in-days":2,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"crossref","award":["F31DK137452"],"award-info":[{"award-number":["F31DK137452"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100005326","name":"Yale University","doi-asserted-by":"crossref","award":["5T32GM007499-43"],"award-info":[{"award-number":["5T32GM007499-43"]}],"id":[{"id":"10.13039\/100005326","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100001680","name":"Charles H. Hood Foundation","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100001680","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000897","name":"Cystic Fibrosis Foundation","doi-asserted-by":"crossref","award":["004741P222"],"award-info":[{"award-number":["004741P222"]}],"id":[{"id":"10.13039\/100000897","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100000057","name":"National Institute of General Medical Sciences","doi-asserted-by":"crossref","award":["1R35GM150645-01"],"award-info":[{"award-number":["1R35GM150645-01"]}],"id":[{"id":"10.13039\/100000057","id-type":"DOI","asserted-by":"crossref"}]},{"DOI":"10.13039\/100005326","name":"Yale University","doi-asserted-by":"crossref","id":[{"id":"10.13039\/100005326","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2025,6,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>The small intestine is well known for its nutrient-absorbing enterocytes; yet equally crucial for homeostasis is a diverse set of secretory cells, all presumed to originate from the same intestinal stem cell. Despite their major roles in intestinal function and health, understanding how the full spectrum of secretory cell types arises remains a longstanding challenge, largely due to their comparative rarity. Here, we investigate the specification of a rare population of small intestinal epithelial cells found in rats and humans but not mice: CFTR high expressers (CHEs). Using pseudotime trajectory analysis of single-cell RNA-sequencing data from rat jejunum, we provide evidence that CHEs are specified along the secretory lineage and appear to employ a second wave of Notch-based signaling to distinguish themselves from other secretory cells. We validate the transcription factors directing these cells from crypt progenitors and demonstrate that Notch signaling is necessary to induce CHE fate in vivo and in vitro. Our findings suggest that Notch reactivation along the secretory lineage specifies CHEs, which may help regulate luminal pH and have direct relevance in cystic fibrosis pathophysiology.<\/jats:p>","DOI":"10.1242\/dev.204277","type":"journal-article","created":{"date-parts":[[2025,5,15]],"date-time":"2025-05-15T06:42:58Z","timestamp":1747291378000},"source":"Crossref","is-referenced-by-count":5,"title":["Evidence of secondary Notch signaling within the rat small intestine"],"prefix":"10.1242","volume":"152","author":[{"given":"Eleanor","family":"Zagoren","sequence":"first","affiliation":[{"name":"Yale School of Medicine 1 Department of Genetics , , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicolas","family":"Dias","sequence":"additional","affiliation":[{"name":"Yale School of Medicine 1 Department of Genetics , , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anderson K.","family":"Santos","sequence":"additional","affiliation":[{"name":"Yale School of Medicine 1 Department of Genetics , , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zachary D.","family":"Smith","sequence":"additional","affiliation":[{"name":"Yale School of Medicine 1 Department of Genetics , , New Haven, CT 06510 ,","place":["USA"]},{"name":"Yale Stem Cell Center, Yale School of Medicine 2 , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nadia A.","family":"Ameen","sequence":"additional","affiliation":[{"name":"Yale School of Medicine 3 Department of Pediatrics\/Gastroenterology and Hepatology , , New Haven, CT 06510 ,","place":["USA"]},{"name":"Yale School of Medicine 4 Department of Cellular and Molecular Physiology , , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1267-7559","authenticated-orcid":false,"given":"Kaelyn","family":"Sumigray","sequence":"additional","affiliation":[{"name":"Yale School of Medicine 1 Department of Genetics , , New Haven, CT 06510 ,","place":["USA"]},{"name":"Yale Stem Cell Center, Yale School of Medicine 2 , New Haven, CT 06510 ,","place":["USA"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"237","published-online":{"date-parts":[[2025,6,3]]},"reference":[{"key":"2025060904311039700_DEV204277C1","doi-asserted-by":"publisher","first-page":"1016","DOI":"10.1016\/0016-5085(95)90198-1","article-title":"A unique subset of rat and human intestinal villus cells express the cystic fibrosis transmembrane conductance regulator","volume":"108","author":"Ameen","year":"1995","journal-title":"Gastroenterology"},{"key":"2025060904311039700_DEV204277C2","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1007\/s004180000164","article-title":"Cellular localization of the cystic fibrosis transmembrane conductance regulator in mouse intestinal tract","volume":"114","author":"Ameen","year":"2000","journal-title":"Histochem. Cell Biol."},{"key":"2025060904311039700_DEV204277C3","doi-asserted-by":"publisher","first-page":"1003","DOI":"10.1038\/nature06196","article-title":"Identification of stem cells in small intestine and colon by marker gene Lgr5","volume":"449","author":"Barker","year":"2007","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C4","doi-asserted-by":"publisher","first-page":"120","DOI":"10.1038\/s41586-022-04552-0","article-title":"Human distal airways contain a multipotent secretory cell that can regenerate alveoli","volume":"604","author":"Basil","year":"2022","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C5","doi-asserted-by":"publisher","first-page":"194","DOI":"10.1016\/j.ydbio.2011.12.009","article-title":"Origin of the brush cell lineage in the mouse intestinal epithelium","volume":"362","author":"Bjerknes","year":"2012","journal-title":"Dev. Biol."},{"key":"2025060904311039700_DEV204277C6","doi-asserted-by":"publisher","first-page":"55159","DOI":"10.3791\/55159-v","article-title":"Forskolin-induced swelling in intestinal organoids: an in vitro assay for assessing drug response in cystic fibrosis patients","volume":"120","author":"Boj","year":"2017","journal-title":"J. Vis. Exp."},{"key":"2025060904311039700_DEV204277C7","doi-asserted-by":"publisher","first-page":"1554","DOI":"10.1016\/j.jcmgh.2022.02.007","article-title":"A proximal-to-distal survey of healthy adult human small intestine and colon epithelium by single-cell transcriptomics","volume":"13","author":"Burclaff","year":"2022","journal-title":"Cell Mol. Gastroenterol. Hepatol."},{"key":"2025060904311039700_DEV204277C8","doi-asserted-by":"publisher","first-page":"108819","DOI":"10.1016\/j.celrep.2021.108819","article-title":"Human gastrointestinal epithelia of the esophagus, stomach, and duodenum resolved at single-cell resolution","volume":"34","author":"Busslinger","year":"2021","journal-title":"Cell Rep."},{"key":"2025060904311039700_DEV204277C9","doi-asserted-by":"publisher","first-page":"98","DOI":"10.1016\/j.ydbio.2015.03.012","article-title":"Notch receptor regulation of intestinal stem cell homeostasis and crypt regeneration","volume":"402","author":"Carulli","year":"2015","journal-title":"Dev. Biol."},{"key":"2025060904311039700_DEV204277C10","doi-asserted-by":"publisher","first-page":"a009753","DOI":"10.1101\/cshperspect.a009753","article-title":"The cystic fibrosis intestine","volume":"3","author":"de Lisle","year":"2013","journal-title":"Cold Spring Harb. Perspect. Med."},{"key":"2025060904311039700_DEV204277C11","doi-asserted-by":"publisher","DOI":"10.1101\/2025.01.24.634747","article-title":"CFTR High Expresser BEST4+ cells are pH-sensing neuropod cells: new implications for intestinal physiology and cystic fibrosis disease","author":"dos Reis","year":"2025","journal-title":"bioRxiv"},{"key":"2025060904311039700_DEV204277C12","doi-asserted-by":"publisher","first-page":"964","DOI":"10.1038\/nature03589","article-title":"Notch signals control the fate of immature progenitor cells in the intestine","volume":"435","author":"Fre","year":"2005","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C13","doi-asserted-by":"publisher","first-page":"2613","DOI":"10.1172\/JCI38662","article-title":"Normal mouse intestinal mucus release requires cystic fibrosis transmembrane regulator-dependent bicarbonate secretion","volume":"119","author":"Garcia","year":"2009","journal-title":"J. Clin. Invest."},{"key":"2025060904311039700_DEV204277C14","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1038\/s41575-018-0081-y","article-title":"Tales from the crypt: new insights into intestinal stem cells","volume":"16","author":"Gehart","year":"2019","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"2025060904311039700_DEV204277C15","doi-asserted-by":"publisher","first-page":"C708","DOI":"10.1152\/ajpcell.00544.2004","article-title":"STa and cGMP stimulate CFTR translocation to the surface of villus enterocytes in rat jejunum and is regulated by protein kinase G","volume":"289","author":"Golin-Bisello","year":"2005","journal-title":"Am. J. Physiol. Cell Physiol."},{"key":"2025060904311039700_DEV204277C16","doi-asserted-by":"publisher","first-page":"1508","DOI":"10.1053\/j.gastro.2018.07.023","article-title":"Sox4 promotes Atoh1-independent intestinal secretory differentiation toward tuft and enteroendocrine fates","volume":"155","author":"Gracz","year":"2018","journal-title":"Gastroenterology"},{"key":"2025060904311039700_DEV204277C17","doi-asserted-by":"publisher","first-page":"333","DOI":"10.1038\/nature24489","article-title":"A single-cell survey of the small intestinal epithelium","volume":"551","author":"Haber","year":"2017","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C18","doi-asserted-by":"publisher","first-page":"840","DOI":"10.1126\/science.1074560","article-title":"Oscillatory expression of the bHLH factor Hes1 regulated by a negative feedback loop","volume":"298","author":"Hirata","year":"2002","journal-title":"Science"},{"key":"2025060904311039700_DEV204277C19","doi-asserted-by":"publisher","first-page":"2023","DOI":"10.1242\/jeb.01611","article-title":"Functional classification of mitochondrion-rich cells in euryhaline Mozambique tilapia (Oreochromis mossambicus) embryos, by means of triple immunofluorescence staining for Na+\/K+-ATPase, Na+\/K+\/2Cl- cotransporter and CFTR anion channel","volume":"208","author":"Hiroi","year":"2005","journal-title":"J. Exp. Biol."},{"key":"2025060904311039700_DEV204277C20","doi-asserted-by":"publisher","first-page":"101524","DOI":"10.1016\/j.xpro.2022.101524","article-title":"Protocol to detect smooth muscle actin-alpha and measure oxidative damage in neonatal mouse intestine","volume":"3","author":"Hu","year":"2022","journal-title":"STAR Protoc."},{"key":"2025060904311039700_DEV204277C21","doi-asserted-by":"publisher","first-page":"G453","DOI":"10.1152\/ajpgi.00094.2013","article-title":"Characterization of CFTR high expresser cells in the intestine","volume":"305","author":"Jakab","year":"2013","journal-title":"Am. J. Physiol. Gastrointest. Liver Physiol."},{"key":"2025060904311039700_DEV204277C22","doi-asserted-by":"publisher","first-page":"2469","DOI":"10.1002\/hep.26254","article-title":"Canonical Notch2 signaling determines biliary cell fates of embryonic hepatoblasts and adult hepatocytes independent of Hes1","volume":"57","author":"Jeliazkova","year":"2013","journal-title":"Hepatology"},{"key":"2025060904311039700_DEV204277C23","doi-asserted-by":"publisher","first-page":"51","DOI":"10.1016\/j.jcmgh.2016.10.001","article-title":"Transcriptional regulation by ATOH1 and its target SPDEF in the Intestine","volume":"3","author":"Lo","year":"2017","journal-title":"Cell Mol. Gastroenterol. Hepatol."},{"key":"2025060904311039700_DEV204277C24","doi-asserted-by":"publisher","first-page":"112520","DOI":"10.1016\/j.celrep.2023.112520","article-title":"Differential cell-cycle control by oscillatory versus sustained Hes1 expression via p21","volume":"42","author":"Maeda","year":"2023","journal-title":"Cell Rep."},{"key":"2025060904311039700_DEV204277C25","doi-asserted-by":"publisher","first-page":"319","DOI":"10.1038\/s41586-018-0393-7","article-title":"A revised airway epithelial hierarchy includes CFTR-expressing ionocytes","volume":"560","author":"Montoro","year":"2018","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C26","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1038\/s42003-022-03013-8","article-title":"Two distinct Notch signals, Delta-like 4\/Notch1 and Jagged-1\/Notch2, antagonistically regulate chemical hepatocarcinogenesis in mice","volume":"5","author":"Nakano","year":"2022","journal-title":"Commun. Biol."},{"key":"2025060904311039700_DEV204277C27","doi-asserted-by":"publisher","first-page":"1275","DOI":"10.1164\/rccm.202008-3198OC","article-title":"Secretory cells dominate airway CFTR expression and function in human airway superficial epithelia","volume":"203","author":"Okuda","year":"2021","journal-title":"Am. J. Respir. Crit. Care. Med."},{"key":"2025060904311039700_DEV204277C28","doi-asserted-by":"publisher","first-page":"49","DOI":"10.1038\/s41586-019-0992-y","article-title":"Colonic epithelial cell diversity in health and inflammatory bowel disease","volume":"567","author":"Parikh","year":"2019","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C29","doi-asserted-by":"publisher","first-page":"1230","DOI":"10.1053\/j.gastro.2011.01.005","article-title":"Dll1- and Dll4-mediated notch signaling are required for homeostasis of intestinal stem cells","volume":"140","author":"Pellegrinet","year":"2011","journal-title":"Gastroenterology"},{"key":"2025060904311039700_DEV204277C30","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1038\/s41586-018-0394-6","article-title":"A single-cell atlas of the airway epithelium reveals the CFTR-rich pulmonary ionocyte","volume":"560","author":"Plasschaert","year":"2018","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C31","doi-asserted-by":"publisher","first-page":"3516","DOI":"10.1038\/s41598-023-30603-1","article-title":"Transcriptomic profile comparison reveals conservation of ionocytes across multiple organs","volume":"13","author":"Pou Casellas","year":"2023","journal-title":"Sci. Rep."},{"key":"2025060904311039700_DEV204277C32","doi-asserted-by":"publisher","first-page":"377","DOI":"10.1038\/embor.2008.7","article-title":"Loss of intestinal crypt progenitor cells owing to inactivation of both Notch1 and Notch2 is accompanied by derepression of CDK inhibitors p27Kip1 and p57Kip2","volume":"9","author":"Riccio","year":"2008","journal-title":"EMBO Rep."},{"key":"2025060904311039700_DEV204277C33","doi-asserted-by":"publisher","first-page":"915","DOI":"10.1038\/ng.165","article-title":"Bmi1 is expressed in vivo in intestinal stem cells","volume":"40","author":"Sangiorgi","year":"2008","journal-title":"Nat. Genet."},{"key":"2025060904311039700_DEV204277C34","doi-asserted-by":"publisher","first-page":"731","DOI":"10.1016\/j.devcel.2020.01.015","article-title":"Jag1 modulates an oscillatory Dll1-Notch-Hes1 signaling module to coordinate growth and fate of pancreatic progenitors","volume":"52","author":"Seymour","year":"2020","journal-title":"Dev. Cell"},{"key":"2025060904311039700_DEV204277C35","doi-asserted-by":"publisher","first-page":"347","DOI":"10.1172\/JCI116966","article-title":"Localization of cystic fibrosis transmembrane conductance regulator mRNA in the human gastrointestinal tract by in situ hybridization","volume":"93","author":"Strong","year":"1994","journal-title":"J. Clin. Invest."},{"key":"2025060904311039700_DEV204277C36","doi-asserted-by":"publisher","first-page":"183","DOI":"10.1016\/j.devcel.2018.03.024","article-title":"Morphogenesis and compartmentalization of the intestinal crypt","volume":"45","author":"Sumigray","year":"2018","journal-title":"Dev. Cell"},{"key":"2025060904311039700_DEV204277C37","doi-asserted-by":"publisher","first-page":"3028","DOI":"10.1016\/j.devcel.2023.11.001","article-title":"Single-cell analysis of shared signatures and transcriptional diversity during zebrafish development","volume":"18","author":"Sur","year":"2023","journal-title":"Dev. Cell."},{"key":"2025060904311039700_DEV204277C38","doi-asserted-by":"publisher","first-page":"1138","DOI":"10.1016\/j.stemcr.2022.03.007","article-title":"Acquisition of NOTCH dependence is a hallmark of human intestinal stem cell maturation","volume":"17","author":"Tsai","year":"2022","journal-title":"Stem Cell Rep."},{"key":"2025060904311039700_DEV204277C39","doi-asserted-by":"publisher","first-page":"1099","DOI":"10.1038\/ncb2581","article-title":"Dll1+ secretory progenitor cells revert to stem cells upon crypt damage","volume":"14","author":"van Es","year":"2012","journal-title":"Nat. Cell Biol."},{"key":"2025060904311039700_DEV204277C40","doi-asserted-by":"publisher","first-page":"598","DOI":"10.1016\/j.stem.2025.02.003","article-title":"Interferon-responsive intestinal BEST4\/CA7(+) cells are targets of bacterial diarrheal toxins","volume":"32","author":"Wang","year":"2025","journal-title":"Cell Stem Cell"},{"key":"2025060904311039700_DEV204277C47","doi-asserted-by":"publisher","DOI":"10.1101\/2024.08.26.609553","article-title":"Notch driven oscillations control cell fate decisions during intestinal homeostasis","author":"Weterings","year":"2024","journal-title":"bioRxiv"},{"key":"2025060904311039700_DEV204277C41","doi-asserted-by":"publisher","first-page":"466","DOI":"10.1073\/pnas.1118857109","article-title":"The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations","volume":"109","author":"Yan","year":"2012","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"2025060904311039700_DEV204277C42","doi-asserted-by":"publisher","first-page":"78","DOI":"10.1016\/j.stem.2017.06.014","article-title":"Intestinal enteroendocrine lineage cells possess homeostatic and injury-inducible stem cell activity","volume":"21","author":"Yan","year":"2017","journal-title":"Cell Stem Cell"},{"key":"2025060904311039700_DEV204277C43","doi-asserted-by":"publisher","first-page":"2155","DOI":"10.1126\/science.1065718","article-title":"Requirement of Math1 for secretory cell lineage commitment in the mouse intestine","volume":"294","author":"Yang","year":"2001","journal-title":"Science"},{"key":"2025060904311039700_DEV204277C44","doi-asserted-by":"publisher","first-page":"857","DOI":"10.1038\/s41586-023-06549-9","article-title":"Transgenic ferret models define pulmonary ionocyte diversity and function","volume":"621","author":"Yuan","year":"2023","journal-title":"Nature"},{"key":"2025060904311039700_DEV204277C45","doi-asserted-by":"publisher","first-page":"65343","DOI":"10.3791\/65343","article-title":"Generation and manipulation of rat intestinal organoids","volume":"196","author":"Zagoren","year":"2023","journal-title":"J. Vis. Exp."},{"key":"2025060904311039700_DEV204277C46","doi-asserted-by":"publisher","first-page":"644","DOI":"10.1242\/dev.113357","article-title":"Enteroendocrine cells are generated from stem cells through a distinct progenitor in the adult Drosophila posterior midgut","volume":"142","author":"Zeng","year":"2015","journal-title":"Development"}],"container-title":["Development"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.204277\/3648581\/dev204277.pdf","content-type":"application\/pdf","content-version":"am","intended-application":"syndication"},{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.204277\/3650915\/dev204277.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/journals.biologists.com\/dev\/article-pdf\/doi\/10.1242\/dev.204277\/3650915\/dev204277.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,6,9]],"date-time":"2025-06-09T04:31:19Z","timestamp":1749443479000},"score":1,"resource":{"primary":{"URL":"https:\/\/journals.biologists.com\/dev\/article\/152\/11\/dev204277\/368176\/Evidence-of-secondary-Notch-signaling-within-the"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,6,1]]},"references-count":47,"journal-issue":{"issue":"11","published-print":{"date-parts":[[2025,6,1]]}},"URL":"https:\/\/doi.org\/10.1242\/dev.204277","relation":{"has-preprint":[{"id-type":"doi","id":"10.1101\/2024.07.15.603542","asserted-by":"object"}]},"ISSN":["0950-1991","1477-9129"],"issn-type":[{"value":"0950-1991","type":"print"},{"value":"1477-9129","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,6,1]]},"article-number":"dev204277"}}