{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,29]],"date-time":"2026-07-29T23:39:01Z","timestamp":1785368341112,"version":"3.55.0"},"reference-count":55,"publisher":"Frontiers Media SA","license":[{"start":{"date-parts":[[2022,9,5]],"date-time":"2022-09-05T00:00:00Z","timestamp":1662336000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001824","name":"Grantov\u00e1 Agentura \u010cesk\u00e9 Republiky","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001824","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001823","name":"Ministerstvo \u0160kolstv\u00ed, Ml\u00e1de\u017ee a T\u011blov\u00fdchovy","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001823","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["frontiersin.org"],"crossmark-restriction":true},"short-container-title":["Front. Cell Dev. Biol."],"abstract":"<jats:p>Leucine Rich Repeat Containing G Protein-Coupled Receptor 5 (LGR5), a Wnt pathway member, has been previously recognised as a stem cell marker in numerous epithelial tissues. In this study, we used <jats:italic>Lgr5-EGFP-CreERT2<\/jats:italic> mice to analyse the distribution of LGR5-positive cells during craniofacial development. LGR5 expressing cells were primarily located in the mesenchyme adjacent to the craniofacial epithelial structures undergoing folding, such as the nasopharyngeal duct, lingual groove, and vomeronasal organ. To follow the fate of LGR5-positive cells, we performed lineage tracing using an inducible Cre knock-in allele in combination with Rosa26-tdTomato reporter mice. The slight expansion of LGR5-positive cells was found around the vomeronasal organ, in the nasal cavity, and around the epithelium in the lingual groove. However, most LGR5 expressing cells remained in their original location, possibly supporting their signalling function for adjacent epithelium rather than exerting their role as progenitor cells for the craniofacial structures. Moreover, <jats:italic>Lgr5<\/jats:italic> knockout mice displayed distinct defects in LGR5-positive areas, especially in the reduction of the nasopharyngeal duct, the alteration of the palatal shelves shape, abnormal epithelial folding in the lingual groove area, and the disruption of salivary gland development. The latter defect manifested as an atypical number and localisation of the glandular ducts. The gene expression of several Wnt pathway members (<jats:italic>Rspo1-3, Axin2<\/jats:italic>) was altered in <jats:italic>Lgr5<\/jats:italic>-deficient animals. However, the difference was not found in sorted EGFP-positive cells obtained from <jats:italic>Lgr5<\/jats:italic><jats:sup><jats:italic>+\/+<\/jats:italic><\/jats:sup> and <jats:italic>Lgr5<\/jats:italic><jats:sup><jats:italic>\u2212\/\u2212<\/jats:italic><\/jats:sup> animals<jats:italic>.<\/jats:italic> Expression profiling of LGR5-positive cells revealed the expression of several markers of mesenchymal cells, antagonists, as well as agonists, of Wnt signalling, and molecules associated with the basal membrane. Therefore, LGR5-positive cells in the craniofacial area represent a very specific population of mesenchymal cells adjacent to the epithelium undergoing folding or groove formation. Our results indicate a possible novel role of LGR5 in the regulation of morphogenetic processes during the formation of complex epithelial structures in the craniofacial areas, a role which is not related to the stem cell properties of LGR5-positive cells as was previously defined for various epithelial tissues.<\/jats:p>","DOI":"10.3389\/fcell.2022.810527","type":"journal-article","created":{"date-parts":[[2022,9,5]],"date-time":"2022-09-05T11:09:01Z","timestamp":1662376141000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":5,"title":["Role of LGR5-positive mesenchymal cells in craniofacial development"],"prefix":"10.3389","volume":"10","author":[{"given":"Krist\u00fdna","family":"Olbertov\u00e1","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Du\u0161an","family":"Hr\u010dkul\u00e1k","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"V\u00edt\u011bzslav","family":"K\u0159\u00ed\u017e","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wojciech","family":"Jesionek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Kubov\u010diak","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Milan","family":"E\u0161ner","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vladim\u00edr","family":"Ko\u0159\u00ednek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcela","family":"Buchtov\u00e1","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1965","published-online":{"date-parts":[[2022,9,5]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1177\/0022034509356401","article-title":"Autosomal-dominant ankyloglossia and tooth number anomalies","volume":"89","author":"Acevedo","year":"2010","journal-title":"J. Dent. Res."},{"key":"B2","doi-asserted-by":"publisher","first-page":"1936","DOI":"10.1038\/s41467-020-15714-x","article-title":"Lgr5+ telocytes are a signaling source at the intestinal villus tip","volume":"11","author":"Bahar Halpern","year":"2020","journal-title":"Nat. Commun."},{"key":"B3","doi-asserted-by":"publisher","first-page":"1681","DOI":"10.1053\/j.gastro.2010.03.002","article-title":"Leucine-rich repeat-containing G-protein-coupled receptors as markers of adult stem cells","volume":"138","author":"Barker","year":"2010","journal-title":"Gastroenterology"},{"key":"B4","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":"B5","doi-asserted-by":"publisher","first-page":"25","DOI":"10.1016\/j.stem.2009.11.013","article-title":"Lgr5+ve stem cells drive self-renewal in the stomach and build long-lived gastric units in vitro","volume":"6","author":"Barker","year":"2010","journal-title":"Cell Stem Cell"},{"key":"B6","doi-asserted-by":"publisher","first-page":"2484","DOI":"10.1242\/dev.083113","article-title":"Lgr proteins in epithelial stem cell biology","volume":"140","author":"Barker","year":"2013","journal-title":"Dev. (Cambridge)"},{"key":"B7","doi-asserted-by":"publisher","first-page":"720","DOI":"10.1002\/stem.2314","article-title":"Lgr5 marks neural crest derived multipotent oral stromal stem cells","volume":"34","author":"Boddupally","year":"2016","journal-title":"Stem Cells"},{"key":"B8","doi-asserted-by":"publisher","first-page":"213","DOI":"10.1111\/j.1365-2818.2006.01706.x","article-title":"A guided tour into subcellular colocalization analysis in light microscopy","volume":"224","author":"Bolte","year":"2006","journal-title":"J. Microsc."},{"key":"B9","doi-asserted-by":"publisher","first-page":"311","DOI":"10.1016\/j.stemcr.2014.01.008","article-title":"Lgr5-positive supporting cells generate new hair cells in the postnatal cochlea","volume":"2","author":"Bramhall","year":"2014","journal-title":"Stem Cell Rep."},{"key":"B10","doi-asserted-by":"publisher","first-page":"11452","DOI":"10.1073\/pnas.1106083108","article-title":"R-spondins function as ligands of the orphan receptors LGR4 and LGR5 to regulate Wnt\/beta-catenin signaling","volume":"108","author":"Carmon","year":"2011","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"B11","doi-asserted-by":"publisher","first-page":"14989","DOI":"10.1074\/jbc.M117.786798","article-title":"LGR5 receptor promotes cell-cell adhesion in stem cells and colon cancer cells via the IQGAP1-Rac1 pathway","volume":"292","author":"Carmon","year":"2017","journal-title":"J. Biol. Chem."},{"key":"B12","doi-asserted-by":"publisher","first-page":"976","DOI":"10.1093\/emboj\/21.5.976","article-title":"The planar polarity gene strabismus regulates convergent extension movements in Xenopus","volume":"21","author":"Darken","year":"2002","journal-title":"EMBO J."},{"key":"B13","doi-asserted-by":"publisher","first-page":"293","DOI":"10.1038\/nature10337","article-title":"Lgr5 homologues associate with Wnt receptors and mediate R-spondin signalling","volume":"476","author":"de Lau","year":"2011","journal-title":"Nature"},{"key":"B14","doi-asserted-by":"publisher","first-page":"300","DOI":"10.1002\/path.4096","article-title":"Developmental stage-specific contribution of LGR5+ cells to basal and luminal epithelial lineages in the postnatal mammary gland","volume":"228","author":"De Visser","year":"2012","journal-title":"J. Pathol."},{"key":"B15","doi-asserted-by":"publisher","first-page":"e23211","DOI":"10.1002\/dvg.23211","article-title":"Salivary gland stem cells: a review of development, regeneration and cancer","volume":"56","author":"Emmerson","year":"2018","journal-title":"Genesis"},{"key":"B16","doi-asserted-by":"publisher","first-page":"276","DOI":"10.1038\/s41587-020-0439-x","article-title":"The nf-core framework for community-curated bioinformatics pipelines","volume":"38","author":"Ewels","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"B17","doi-asserted-by":"publisher","first-page":"17934","DOI":"10.1038\/s41598-019-54216-9","article-title":"Fractionated head and neck irradiation impacts taste progenitors, differentiated taste cells, and Wnt\/\u03b2-catenin signaling in adult mice","volume":"9","author":"Gaillard","year":"2019","journal-title":"Sci. Rep."},{"key":"B19","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1038\/embor.2011.175","article-title":"LGR4 and LGR5 are R-spondin receptors mediating Wnt\/\u03b2-catenin and Wnt\/PCP signalling","volume":"12","author":"Glinka","year":"2011","journal-title":"EMBO Rep."},{"key":"B20","doi-asserted-by":"publisher","first-page":"715","DOI":"10.2353\/ajpath.2009.080758","article-title":"Wnt signaling, Lgr5, and stem cells in the intestine and skin","volume":"174","author":"Haegebarth","year":"2009","journal-title":"Am. J. Pathol."},{"key":"B21","doi-asserted-by":"publisher","first-page":"195","DOI":"10.1038\/nature11019","article-title":"ZNRF3 promotes Wnt receptor turnover in an R-spondin-sensitive manner","volume":"485","author":"Hao","year":"2012","journal-title":"Nature"},{"key":"B22","doi-asserted-by":"publisher","first-page":"273","DOI":"10.1006\/bbrc.1998.9882","article-title":"Identification of a novel seven-transmembrane receptor with homology to glycoprotein receptors and its expression in the adult and developing mouse","volume":"254","author":"Hermey","year":"1999","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"B56","doi-asserted-by":"publisher","first-page":"884","DOI":"10.1093\/nar\/gkaa942","volume":"49","author":"Howe","year":"2021","journal-title":"Nucleic Acids Res"},{"key":"B23","doi-asserted-by":"publisher","first-page":"1172","DOI":"10.1128\/MCB.22.4.1172-1183.2002","article-title":"Wnt\/beta-catenin\/Tcf signaling induces the transcription of Axin2, a negative regulator of the signaling pathway","volume":"22","author":"Jho","year":"2002","journal-title":"Mol. Cell. Biol."},{"key":"B24","doi-asserted-by":"publisher","first-page":"844","DOI":"10.1002\/dvdy.24124","article-title":"R-spondins\/Lgrs expression in tooth development","volume":"243","author":"Kawasaki","year":"2014","journal-title":"Dev. Dyn."},{"key":"B25","doi-asserted-by":"publisher","first-page":"777","DOI":"10.1016\/s0898-6568(99)00054-6","article-title":"Roles of Axin in the Wnt signalling pathway","volume":"11","author":"Kikuchi","year":"1999","journal-title":"Cell. Signal."},{"key":"B26","doi-asserted-by":"publisher","first-page":"463","DOI":"10.1006\/cbir.2002.0901","article-title":"Direct evidence for a role of beta-catenin\/LEF-1 signaling pathway in induction of EMT","volume":"26","author":"Kim","year":"2002","journal-title":"Cell Biol. Int."},{"key":"B27","doi-asserted-by":"publisher","first-page":"289","DOI":"10.1053\/j.gastro.2014.05.007","article-title":"Stem cells marked by the R-spondin receptor LGR5","volume":"147","author":"Koo","year":"2014","journal-title":"Gastroenterology"},{"key":"B28","doi-asserted-by":"publisher","first-page":"665","DOI":"10.1038\/nature11308","article-title":"Tumour suppressor RNF43 is a stem-cell E3 ligase that induces endocytosis of Wnt receptors","volume":"488","author":"Koo","year":"2012","journal-title":"Nature"},{"key":"B29","doi-asserted-by":"publisher","first-page":"20","DOI":"10.3390\/genes9010020","article-title":"Wnt, RSPO and hippo signalling in the intestine and intestinal stem cells","volume":"9","author":"Kriz","year":"2018","journal-title":"Genes (Basel)"},{"key":"B30","doi-asserted-by":"publisher","first-page":"551","DOI":"10.1002\/pro.2446","article-title":"Structure and function of LGR5: an enigmatic G-protein coupled receptor marking stem cells","volume":"23","author":"Kumar","year":"2014","journal-title":"Protein Sci."},{"key":"B31","doi-asserted-by":"publisher","first-page":"1149","DOI":"10.1016\/j.cell.2017.07.028","article-title":"Anatomically and functionally distinct lung mesenchymal populations marked by Lgr5 and Lgr6","volume":"170","author":"Lee","year":"2017","journal-title":"Cell"},{"key":"B32","doi-asserted-by":"publisher","first-page":"1614","DOI":"10.1002\/ajmg.a.38690","article-title":"Familial autosomal dominant severe ankyloglossia with tooth abnormalities","volume":"176","author":"Lenormand","year":"2018","journal-title":"Am. J. Med. Genet. A"},{"key":"B33","doi-asserted-by":"publisher","first-page":"108535","DOI":"10.1016\/j.celrep.2020.108535","article-title":"Lgr5 marks adult progenitor cells contributing to skeletal muscle regeneration and sarcoma formation","volume":"33","author":"Leung","year":"2020","journal-title":"Cell Rep."},{"key":"B34","doi-asserted-by":"publisher","first-page":"919","DOI":"10.1083\/jcb.136.4.919","article-title":"Cross-talk between adherens junctions and desmosomes depends on plakoglobin","volume":"136","author":"Lewis","year":"1997","journal-title":"J. Cell Biol."},{"key":"B35","doi-asserted-by":"publisher","first-page":"359","DOI":"10.1083\/jcb.200403050","article-title":"Beta-catenin is required for endothelial-mesenchymal transformation during heart cushion development in the mouse","volume":"166","author":"Liebner","year":"2004","journal-title":"J. Cell Biol."},{"key":"B36","doi-asserted-by":"publisher","first-page":"550","DOI":"10.1186\/s13059-014-0550-8","article-title":"Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2","volume":"15","author":"Love","year":"2014","journal-title":"Genome Biol."},{"key":"B37","doi-asserted-by":"publisher","first-page":"10106","DOI":"10.1038\/srep10106","article-title":"Characterization of a self-renewing and multi-potent cell population isolated from human minor salivary glands","volume":"5","author":"Lu","year":"2015","journal-title":"Sci. Rep."},{"key":"B38","doi-asserted-by":"publisher","first-page":"539","DOI":"10.1038\/nm.4076","article-title":"LGR4 is a receptor for RANKL and negatively regulates osteoclast differentiation and bone resorption","volume":"22","author":"Luo","year":"2016","journal-title":"Nat. Med."},{"key":"B39","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/nn.2467","article-title":"A robust and high-throughput Cre reporting and characterization system for the whole mouse brain","volume":"13","author":"Madisen","year":"2010","journal-title":"Nat. Neurosci."},{"key":"B40","doi-asserted-by":"publisher","first-page":"416","DOI":"10.1002\/dvdy.20135","article-title":"Beta-catenin signaling marks the prospective site of primitive streak formation in the mouse embryo","volume":"231","author":"Mohamed","year":"2004","journal-title":"Dev. Dyn."},{"key":"B41","doi-asserted-by":"publisher","first-page":"9736","DOI":"10.1128\/MCB.24.22.9736-9743.2004","article-title":"Neonatal lethality of LGR5 null mice is associated with ankyloglossia and gastrointestinal distension","volume":"24","author":"Morita","year":"2004","journal-title":"Mol. Cell. Biol."},{"key":"B42","doi-asserted-by":"publisher","first-page":"80","DOI":"10.1016\/j.jdsr.2019.03.001","article-title":"R-spondin signaling as a pivotal regulator of tissue development and homeostasis","volume":"55","author":"Nagano","year":"2019","journal-title":"Jpn. Dent. Sci. Rev."},{"key":"B43","doi-asserted-by":"publisher","first-page":"1291","DOI":"10.1083\/jcb.200306024","article-title":"TGFbeta3 signaling activates transcription of the LEF1 gene to induce epithelial mesenchymal transformation during mouse palate development","volume":"163","author":"Nawshad","year":"2003","journal-title":"J. Cell Biol."},{"key":"B44","doi-asserted-by":"publisher","first-page":"675","DOI":"10.1016\/j.archoralbio.2004.05.007","article-title":"Transforming growth factor beta (TGFbeta) signalling in palatal growth, apoptosis and epithelial mesenchymal transformation (EMT)","volume":"49","author":"Nawshad","year":"2004","journal-title":"Arch. Oral Biol."},{"key":"B45","doi-asserted-by":"publisher","first-page":"119195","DOI":"10.1016\/j.gep.2021.119195","article-title":"Expression of R-spondins\/Lgrs in development of movable craniofacial organs","volume":"41","author":"Nihara","year":"2021","journal-title":"Gene Expr. Patterns."},{"key":"B46","unstructured":"Wnt target genes\n            NusseR.\n          2022"},{"key":"B47","doi-asserted-by":"publisher","first-page":"417","DOI":"10.1038\/nmeth.4197","article-title":"Salmon provides fast and bias-aware quantification of transcript expression","volume":"14","author":"Patro","year":"2017","journal-title":"Nat. Methods"},{"key":"B48","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1038\/nmeth.2089","article-title":"NIH image to ImageJ: 25 years of image analysis","volume":"9","author":"Schneider","year":"2012","journal-title":"Nat. Methods"},{"key":"B49","doi-asserted-by":"publisher","first-page":"433","DOI":"10.1186\/s12859-021-04344-9","article-title":"CellProfiler 4: improvements in speed, utility and usability","volume":"22","author":"Stirling","year":"2021","journal-title":"BMC Bioinforma."},{"key":"B50","doi-asserted-by":"publisher","first-page":"564","DOI":"10.1038\/s41586-018-0118-y","article-title":"RSPO2 inhibition of RNF43 and ZNRF3 governs limb development independently of LGR4\/5\/6","volume":"557","author":"Szenker-Ravi","year":"2018","journal-title":"Nature"},{"key":"B51","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/sj.onc.1204052","article-title":"Inhibition of integrin linked kinase (ILK) suppresses beta-catenin-Lef\/Tcf-dependent transcription and expression of the E-cadherin repressor, snail, in APC-\/- human colon carcinoma cells","volume":"20","author":"Tan","year":"2001","journal-title":"Oncogene"},{"key":"B52","doi-asserted-by":"publisher","first-page":"81","DOI":"10.1038\/nature13851","article-title":"Identification of multipotent mammary stem cells by protein C receptor expression","volume":"517","author":"Wang","year":"2015","journal-title":"Nature"},{"key":"B53","doi-asserted-by":"publisher","first-page":"616","DOI":"10.1002\/dvdy.22564","article-title":"The planar cell polarity pathway in vertebrate development","volume":"240","author":"Wansleeben","year":"2011","journal-title":"Dev. Dyn."},{"key":"B54","doi-asserted-by":"publisher","first-page":"59","DOI":"10.1146\/annurev.cellbio.14.1.59","article-title":"Mechanisms of Wnt signaling in development","volume":"14","author":"Wodarz","year":"1998","journal-title":"Annu. Rev. Cell Dev. Biol."},{"key":"B55","doi-asserted-by":"publisher","first-page":"992","DOI":"10.1002\/stem.1338","article-title":"Lgr5-EGFP marks taste bud stem\/progenitor cells in posterior tongue","volume":"31","author":"Yee","year":"2013","journal-title":"Stem Cells"}],"container-title":["Frontiers in Cell and Developmental Biology"],"original-title":[],"link":[{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2022.810527\/full","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,9,5]],"date-time":"2022-09-05T11:09:07Z","timestamp":1662376147000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.frontiersin.org\/articles\/10.3389\/fcell.2022.810527\/full"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,5]]},"references-count":55,"alternative-id":["10.3389\/fcell.2022.810527"],"URL":"https:\/\/doi.org\/10.3389\/fcell.2022.810527","relation":{},"ISSN":["2296-634X"],"issn-type":[{"value":"2296-634X","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,5]]},"article-number":"810527"}}