{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,10]],"date-time":"2026-07-10T02:34:39Z","timestamp":1783650879774,"version":"3.55.0"},"reference-count":41,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T00:00:00Z","timestamp":1715299200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/publicdomain\/zero\/1.0\/"}],"funder":[{"name":"the Community Program in Life Sciences from the Ministry of Education, Culture, Sports, Science and Technology of Japan","award":["20K10191"],"award-info":[{"award-number":["20K10191"]}]},{"name":"the Community Program in Life Sciences from the Ministry of Education, Culture, Sports, Science and Technology of Japan","award":["20K09895"],"award-info":[{"award-number":["20K09895"]}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Entheses are classified into three types: fibrocartilaginous, fibrous, and periosteal insertions. However, the mechanism behind the development of fibrous entheses and periosteal insertions remains unclear. Since both entheses are part of the temporomandibular joint (TMJ), this study analyzes the TMJ entheses. Here, we show that SOX9 expression is negatively regulated during TMJ enthesis development, unlike fibrocartilage entheses which are modularly formed by SCX and SOX9 positive progenitors. The TMJ entheses was adjacent to the intramembranous bone rather than cartilage. SOX9 expression was diminished during TMJ enthesis development. To clarify the functional role of <jats:italic>Sox9<\/jats:italic> in the development of TMJ entheses, we examined these structures in TMJ using <jats:italic>Wnt1Cre;Sox9<\/jats:italic><jats:sup><jats:italic>flox\/+<\/jats:italic><\/jats:sup> reporter mice. <jats:italic>Wnt1Cre;Sox9<\/jats:italic><jats:sup><jats:italic>flox\/+<\/jats:italic><\/jats:sup> mice showed enthesial deformation at the TMJ. Next, we also observed a diminished SOX9 expression area at the enthesis in contact with the clavicle\u2019s membranous bone portion, similar to the TMJ entheses. Together, these findings reveal that the timing of SOX9 expression varies with the ossification development mode.<\/jats:p>","DOI":"10.1371\/journal.pone.0301080","type":"journal-article","created":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T17:38:34Z","timestamp":1715362714000},"page":"e0301080","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["Downregulation of SOX9 expression in developing entheses adjacent to intramembranous bone"],"prefix":"10.1371","volume":"19","author":[{"given":"Asahi","family":"Kitamura","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9683-6678","authenticated-orcid":true,"given":"Masahito","family":"Yamamoto","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hidetomo","family":"Hirouchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Genji","family":"Watanabe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuichiro","family":"Taniguchi","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sayo","family":"Sekiya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satoshi","family":"Ishizuka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juhee","family":"Jeong","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kazunari","family":"Higa","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shuichiro","family":"Yamashita","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Shinichi","family":"Abe","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"340","published-online":{"date-parts":[[2024,5,10]]},"reference":[{"key":"pone.0301080.ref001","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1016\/S0074-7696(00)96003-0","article-title":"The cell and developmental biology of tendons and ligaments","volume":"196","author":"M Benjamin","year":"2000","journal-title":"Int Rev Cytol"},{"key":"pone.0301080.ref002","first-page":"151","article-title":"Entheses\u2014the bony attachments of tendons and ligaments","volume":"106","author":"M Benjamin","year":"2001","journal-title":"Ital J Anat Embryol"},{"key":"pone.0301080.ref003","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1007\/s12565-019-00523-0","article-title":"Mechanism of muscle-tendon-bone complex development in the head","volume":"95","author":"M Yamamoto","year":"2020","journal-title":"Anat Sci Int"},{"key":"pone.0301080.ref004","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/8756-3282(96)00116-0","article-title":"Adaptive changes in trabecular architecture in relation to functional strain patterns and disuse","volume":"19","author":"AA Biewener","year":"1996","journal-title":"Bone"},{"key":"pone.0301080.ref005","doi-asserted-by":"crossref","first-page":"931","DOI":"10.1016\/S1095-6433(02)00138-1","article-title":"The skeletal attachment of tendons\u2014tendon \u201centheses\u201d.","volume":"133","author":"M Benjamin","year":"2002","journal-title":"Comp Biochem Physiol A Mol Integr Physiol"},{"key":"pone.0301080.ref006","first-page":"35","article-title":"The development and morphogenesis of the tendon-to-bone insertion\u2013what development can teach us about healing","volume":"10","author":"S Thomopoulos","year":"2010","journal-title":"J. Musculoskelet. Neuronal Interact"},{"key":"pone.0301080.ref007","doi-asserted-by":"crossref","first-page":"1039","DOI":"10.1089\/ten.tea.2009.0499","article-title":"Fibrocartilage tissue engineering: the role of the stress environment on cell morphology and matrix expression","volume":"17","author":"S Thomopoulos","year":"2011","journal-title":"Tissue Eng"},{"key":"pone.0301080.ref008","doi-asserted-by":"crossref","first-page":"3855","DOI":"10.1242\/dev.128.19.3855","article-title":"Analysis of the tendon cell fate using Scleraxis, a specific marker for tendons and ligaments","volume":"128","author":"R Schweitzer","year":"2001","journal-title":"Development"},{"key":"pone.0301080.ref009","doi-asserted-by":"crossref","first-page":"1677","DOI":"10.1002\/dvdy.21179","article-title":"Generation of transgenic tendon reporters, ScxGFP and ScxAP, using regulatory elements of the scleraxis gene","volume":"236","author":"BA Pryce","year":"2007","journal-title":"Dev. Dyn"},{"issue":"14","key":"pone.0301080.ref010","doi-asserted-by":"crossref","first-page":"2697","DOI":"10.1242\/dev.001933","article-title":"Regulation of tendon differentiation by scleraxis distinguishes force-transmitting tendons from muscle-anchoring tendons","volume":"134","author":"ND Murchison","year":"2007","journal-title":"Development"},{"key":"pone.0301080.ref011","first-page":"861","article-title":"Bone ridge patterning during musculoskeletal assembly is mediated through SCX regulation of Bmp4 at the tendon-skeleton junction.","volume":"17","author":"E Blitz","year":"2009","journal-title":"Dev"},{"issue":"13","key":"pone.0301080.ref012","doi-asserted-by":"crossref","first-page":"2680","DOI":"10.1242\/dev.093906","article-title":"Tendon-bone attachment unit is formed modularly by a distinct pool of Scx- and Sox9-positive progenitors","volume":"140","author":"E Blitz","year":"2013","journal-title":"Development"},{"issue":"11","key":"pone.0301080.ref013","doi-asserted-by":"crossref","first-page":"2280","DOI":"10.1242\/dev.096354","article-title":"Scx+\/Sox9+ progenitors contribute to the establishment of the junction between cartilage and tendon\/ligament","volume":"140","author":"Y Sugimoto","year":"2013","journal-title":"Development"},{"issue":"1","key":"pone.0301080.ref014","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1242\/dev.112714","article-title":"Enthesis fibrocartilage cells originate from a population of Hedgehog-responsive cells modulated by the loading environment","volume":"142","author":"AG Schwartz","year":"2015","journal-title":"Development"},{"issue":"2","key":"pone.0301080.ref015","first-page":"77","article-title":"Thickness of the soft tissue layers and the articular disk in the temporomandibular joint.","volume":"35","author":"T Hansson","year":"2009","journal-title":"Acta Odontologica Scandinavica."},{"issue":"6","key":"pone.0301080.ref016","doi-asserted-by":"crossref","first-page":"599","DOI":"10.1016\/j.archoralbio.2011.10.004","article-title":"The attachments of the temporomandibular joint disc: A biochemical and histological investigation","volume":"57","author":"VP Willard","year":"2012","journal-title":"Archives of Oral Biology"},{"issue":"4","key":"pone.0301080.ref017","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1111\/j.1469-7580.2012.01478.x","article-title":"Fetal developmental change in topographical relationship between the human lateral pterygoid muscle and buccal nerve","volume":"220","author":"Y Katori","year":"2012","journal-title":"J Anat"},{"issue":"2","key":"pone.0301080.ref018","doi-asserted-by":"crossref","first-page":"153","DOI":"10.5603\/FM.2014.0023","article-title":"Individual variations in the vascular content of retrodiscal tissue in the temporomandibular joint: a study using histological sections of human foetuses and magnetic resonance images of adults without pathology.","volume":"73","author":"M Yamamoto","year":"2014","journal-title":"Folia Morphol (Warsz)."},{"key":"pone.0301080.ref019","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1007\/s004290000107","article-title":"Tendon entheses of the human masticatory muscles","volume":"202","author":"T Hems","year":"2000","journal-title":"Anat Embryol"},{"key":"pone.0301080.ref020","doi-asserted-by":"crossref","first-page":"8425","DOI":"10.1038\/s41598-020-65339-9","article-title":"Switching of Sox9 expression during musculoskeletal system development","volume":"10","author":"R Nagakura","year":"2020","journal-title":"Sci Rep"},{"key":"pone.0301080.ref021","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1016\/j.gep.2016.03.001","article-title":"Expression of forkhead box transcription factor genes Foxp1 and Foxp2 during jaw development","volume":"20","author":"JM Cesario","year":"2016","journal-title":"Gene Expr Patterns"},{"key":"pone.0301080.ref022","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1007\/BF00267823","article-title":"Single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ hybridization using digoxigenin-labelled cRNA probes","volume":"100","author":"N Schaeren-Wiemers","year":"1993","journal-title":"Histochemistry"},{"key":"pone.0301080.ref023","doi-asserted-by":"crossref","first-page":"919","DOI":"10.1016\/j.archoralbio.2010.07.010","article-title":"Effects of restriction of fetal jaw movement on prenatal development of the temporalis muscle","volume":"55","author":"E Jahan","year":"2010","journal-title":"Arch Oral Biol"},{"key":"pone.0301080.ref024","doi-asserted-by":"crossref","first-page":"871","DOI":"10.1016\/j.archoralbio.2014.03.011","article-title":"Desmin and nerve terminal expression during embryonic development of the lateral pterygoid muscle in mice","volume":"59","author":"M Yamamoto","year":"2014","journal-title":"Arch Oral Biol"},{"key":"pone.0301080.ref025","doi-asserted-by":"crossref","first-page":"dev170241","DOI":"10.1242\/dev.170241","article-title":"FGF signaling patterns cell fate at the interface between tendon and bone","volume":"146","author":"RR Roberts","year":"2019","journal-title":"Development"},{"key":"pone.0301080.ref026","doi-asserted-by":"crossref","first-page":"1028","DOI":"10.1016\/j.bbrc.2019.01.059","article-title":"Runx2 function in cells of neural crest origin during intramembranous ossification","volume":"509","author":"Y Shirai","year":"2019","journal-title":"Biochem Biophys Res Commun"},{"key":"pone.0301080.ref027","volume-title":"Gray\u2019s anatomy (42nd ed.)","author":"S. Standring","year":"2020"},{"key":"pone.0301080.ref028","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1016\/j.archoralbio.2017.06.001","article-title":"Developmental mechanism of muscle-tendon-bone complex in the fetal soft palate","volume":"82","author":"M Nara","year":"2017","journal-title":"Arch Oral Biol"},{"key":"pone.0301080.ref029","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.archoralbio.2017.12.027","article-title":"Developmental characteristics of secondary cartilage in the mandibular condyle and sphenoid bone in mice","volume":"89","author":"H Hirouchi","year":"2018","journal-title":"Arch Oral Biol"},{"key":"pone.0301080.ref030","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/0003-9969(95)00105-0","article-title":"A histological study of the developing condylar cartilage of the fetal mouse mandible using coronal sections","volume":"41","author":"S Shibata","year":"1996","journal-title":"Arch Oral Biol"},{"key":"pone.0301080.ref031","doi-asserted-by":"crossref","first-page":"561","DOI":"10.1046\/j.1469-7580.1997.19140561.x","article-title":"Immunohistochemistry of collagen types II and X, and enzyme-histochemistry of alkaline phosphatase in the developing condylar cartilage of the fetal mouse mandible","volume":"191","author":"S Shibata","year":"1997","journal-title":"J Anat"},{"issue":"8","key":"pone.0301080.ref032","doi-asserted-by":"crossref","first-page":"1872","DOI":"10.1002\/jor.25210","article-title":"The effect of mechanical stress on enthesis homeostasis in a rat Achilles enthesis organ culture model","volume":"40","author":"T Saito","year":"2022","journal-title":"J Orthop Res"},{"issue":"1","key":"pone.0301080.ref033","doi-asserted-by":"crossref","first-page":"e0189668","DOI":"10.1371\/journal.pone.0189668","article-title":"Biomarkers for tissue engineering of the tendon-bone interface","volume":"13","author":"LA Kuntz","year":"2018","journal-title":"PLoS One"},{"key":"pone.0301080.ref034","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.3390\/ijms20071694","article-title":"Regulation of proliferation, differentiation and functions of osteoblasts by Runx2","volume":"20","author":"T. Komori","year":"2019","journal-title":"Int J Mol Sci."},{"key":"pone.0301080.ref035","doi-asserted-by":"crossref","first-page":"333","DOI":"10.32098\/mltj.03.2014.12","article-title":"The enthesis: a review of the tendon-to-bone insertion.","volume":"4","author":"J Apostolakos","year":"2014","journal-title":"Muscles Ligaments Tendons J"},{"key":"pone.0301080.ref036","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1146\/annurev-bioeng-071910-124656","article-title":"Functional attachment of soft tissues to bone: development, healing, and tissue engineering","volume":"15","author":"HH Lu","year":"2013","journal-title":"Annu Rev Biomed Eng"},{"key":"pone.0301080.ref037","doi-asserted-by":"crossref","first-page":"410","DOI":"10.1111\/dgd.12136","article-title":"Growth and differentiation of a long bone in limb development, repair and regeneration","volume":"56","author":"S Egawa","year":"2014","journal-title":"Dev Growth Differ"},{"key":"pone.0301080.ref038","doi-asserted-by":"crossref","first-page":"e0242286","DOI":"10.1371\/journal.pone.0242286","article-title":"Role of Scx+\/Sox9+ cells as potential progenitor cells for postnatal supraspinatus enthesis formation and healing after injury in mice.","volume":"15","author":"K Ideo","year":"2020","journal-title":"PLoS One"},{"issue":"1","key":"pone.0301080.ref039","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1359\/JBMR.051005","article-title":"Initial characterization of PTH-related protein gene-driven lacZ expression in the mouse","volume":"21","author":"X Chen","year":"2006","journal-title":"J Bone Miner Res"},{"issue":"5","key":"pone.0301080.ref040","doi-asserted-by":"crossref","first-page":"752","DOI":"10.1016\/j.bone.2007.07.020","article-title":"Mechanical regulation of PTHrP expression in entheses","volume":"41","author":"X Chen","year":"2007","journal-title":"Bone"},{"key":"pone.0301080.ref041","doi-asserted-by":"crossref","first-page":"598","DOI":"10.1002\/jbmr.1801","article-title":"PTHrP regulates the modeling of cortical bone surfaces at fibrous insertion sites during growth","volume":"28","author":"M Wang","year":"2013","journal-title":"J Bone Miner Res"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0301080","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,10]],"date-time":"2024-05-10T17:38:53Z","timestamp":1715362733000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0301080"}},"subtitle":[],"editor":[{"given":"Gianpaolo","family":"Papaccio","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"editor"}]}],"short-title":[],"issued":{"date-parts":[[2024,5,10]]},"references-count":41,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5,10]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0301080","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,10]]}}}