{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,21]],"date-time":"2026-01-21T12:23:52Z","timestamp":1768998232976,"version":"3.49.0"},"reference-count":24,"publisher":"Oxford University Press (OUP)","issue":"12","license":[{"start":{"date-parts":[[2015,6,19]],"date-time":"2015-06-19T00:00:00Z","timestamp":1434672000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/journals\/pages\/open_access\/funder_policies\/chorus\/standard_publication_model"}],"funder":[{"name":"Funds from HSDM (to BL), Postdoctoral Fellowships for Research Abroad from Japan Society for the Promotion of Science (to KA)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2015,12,1]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n               <jats:sec>\n                  <jats:title>\u2003<\/jats:title>\n                  <jats:p>Indian hedgehog (Ihh) is widely recognized as an essential factor for proper skeletal development. Previous in vivo studies using mutant Ihh mouse models were limited by perinatal lethality or carried out after a growth plate formed. Thus the important role of Ihh in mesenchymal cell differentiation has not been investigated. In this study, we established Prx1-Cre;Ihhfl\/fl mice to ablate Ihh specifically in limb mesenchyme to allow us to observe the phenotype continuously from prenatal development to 3 weeks of age. Mutant mice displayed severe limb abnormalities characterized by complete lack of secondary ossification center and growth plate, indicating an essential role for Ihh in the development of these structures. Interestingly, we discovered that osteoblast differentiation and bone formation could occur in conditions of deficient Ihh. This is a novel finding that has not been observed because of the early lethality of previous Ihh mutants. Mature osteoblasts expressing osteocalcin could be detected in the center of mutant bones at postnatal day 10 (P10). Osteoclasts and blood vessel formation were also present, suggesting active bone remodeling. Histomorphometric analyses show a significant increase in osteoclast number with no major changes in bone formation rate at 3 weeks of age. Mutant long bones in the limbs were deformed, with cortices comprised of irregular woven bone. Also, there was a marked decrease in gene expression of osteoblastic and osteocytic markers. Moreover, mutant long bones displayed bone dysplasia in which we observed increased osteoclast activity and partially reduced osteoblastic and osteocytic differentiation that lead ultimately to loss of bone structures at 3 weeks of age. In summary, our data show for the first time, the presence of mature osteoblasts in long bones of the limbs despite the complete loss of growth plate formation due to Ihh deficiency. These data indicate an important function for Ihh in regulating limb mesenchymal cell differentiation. \u00a9 2015 American Society for Bone and Mineral Research.<\/jats:p>\n               <\/jats:sec>","DOI":"10.1002\/jbmr.2582","type":"journal-article","created":{"date-parts":[[2015,6,20]],"date-time":"2015-06-20T08:41:23Z","timestamp":1434789683000},"page":"2262-2272","source":"Crossref","is-referenced-by-count":37,"title":["Conditional Deletion of Indian Hedgehog in Limb Mesenchyme Results in Complete Loss of Growth Plate Formation but Allows Mature Osteoblast Differentiation"],"prefix":"10.1093","volume":"30","author":[{"given":"Katsuhiko","family":"Amano","sequence":"first","affiliation":[{"name":"Department of Oral Medicine, Infection, and ImmunityHarvard School of Dental MedicineBostonMAUSA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael J","family":"Densmore","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine, Infection, and ImmunityHarvard School of Dental MedicineBostonMAUSA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Beate","family":"Lanske","sequence":"additional","affiliation":[{"name":"Department of Oral Medicine, Infection, and ImmunityHarvard School of Dental MedicineBostonMAUSA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2015,6,19]]},"reference":[{"issue":"6","key":"2024020319001019700_jbmr2582-bib-0001","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1016\/S0955-0674(00)00276-3","article-title":"Regulatory mechanisms in the pathways of cartilage and bone formation","volume":"13","author":"de Crombrugghe","year":"2001","journal-title":"Curr Opin Cell Biol"},{"issue":"6937","key":"2024020319001019700_jbmr2582-bib-0002","doi-asserted-by":"crossref","first-page":"332","DOI":"10.1038\/nature01657","article-title":"Developmental regulation of the growth plate","volume":"423","author":"Kronenberg","year":"2003","journal-title":"Nature"},{"issue":"4","key":"2024020319001019700_jbmr2582-bib-0003","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1038\/ng577","article-title":"Mutations in IHH, encoding Indian hedgehog, cause brachydactyly type A-1","volume":"28","author":"Gao","year":"2001","journal-title":"Nat Genet"},{"issue":"4","key":"2024020319001019700_jbmr2582-bib-0004","doi-asserted-by":"crossref","first-page":"1040","DOI":"10.1086\/374318","article-title":"Homozygous mutations in IHH cause acrocapitofemoral dysplasia, an autosomal recessive disorder with cone-shaped epiphyses in hands and hips","volume":"72","author":"Hellemans","year":"2003","journal-title":"Am J Hum Genet"},{"issue":"16","key":"2024020319001019700_jbmr2582-bib-0005","doi-asserted-by":"crossref","first-page":"2072","DOI":"10.1101\/gad.13.16.2072","article-title":"Indian hedgehog signaling regulates proliferation and differentiation of chondrocytes and is essential for bone formation","volume":"13","author":"St-Jacques","year":"1999","journal-title":"Genes Dev"},{"issue":"4","key":"2024020319001019700_jbmr2582-bib-0006","doi-asserted-by":"crossref","first-page":"453","DOI":"10.1002\/path.1870","article-title":"Conditional deletion of Indian hedgehog from collagen type 2alpha1-expressing cells results in abnormal endochondral bone formation","volume":"207","author":"Razzaque","year":"2005","journal-title":"J Pathol"},{"issue":"15","key":"2024020319001019700_jbmr2582-bib-0007","doi-asserted-by":"crossref","first-page":"6382","DOI":"10.1073\/pnas.0608449104","article-title":"Indian Hedgehog produced by postnatal chondrocytes is essential for maintaining a growth plate and trabecular bone","volume":"104","author":"Maeda","year":"2007","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"6","key":"2024020319001019700_jbmr2582-bib-0008","doi-asserted-by":"crossref","first-page":"1309","DOI":"10.1242\/dev.01006","article-title":"Ihh signaling is directly required for the osteoblast lineage in the endochondral skeleton","volume":"131","author":"Long","year":"2004","journal-title":"Development"},{"issue":"2","key":"2024020319001019700_jbmr2582-bib-0009","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1002\/gene.10092","article-title":"Expression of Cre Recombinase in the developing mouse limb bud driven by a Prxl enhancer","volume":"33","author":"Logan","year":"2002","journal-title":"Genesis"},{"issue":"5","key":"2024020319001019700_jbmr2582-bib-0010","doi-asserted-by":"crossref","first-page":"284","DOI":"10.1007\/BF02556360","article-title":"Bone histomorphometry: proposed system for standardization of nomenclature, symbols, and units","volume":"42","author":"Parfitt","year":"1988","journal-title":"Calcif Tissue Int"},{"issue":"6","key":"2024020319001019700_jbmr2582-bib-0011","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1002\/jbmr.5650020617","article-title":"Bone histomorphometry: standardization of nomenclature, symbols, and units. Report of the ASBMR Histomorphometry Nomenclature Committee","volume":"2","author":"Parfitt","year":"1987","journal-title":"J Bone Miner Res"},{"issue":"1","key":"2024020319001019700_jbmr2582-bib-0012","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1172\/JCI65910","article-title":"Estrogen receptor-alpha signaling in osteoblast progenitors stimulates cortical bone accrual","volume":"123","author":"Almeida","year":"2013","journal-title":"J Clin Invest"},{"issue":"4","key":"2024020319001019700_jbmr2582-bib-0013","doi-asserted-by":"crossref","first-page":"390","DOI":"10.1007\/s00774-013-0429-x","article-title":"Periosteal cells are a major source of soft callus in bone fracture","volume":"31","author":"Murao","year":"2013","journal-title":"J Bone Miner Metab"},{"issue":"5","key":"2024020319001019700_jbmr2582-bib-0014","doi-asserted-by":"crossref","first-page":"1057","DOI":"10.1242\/dev.01649","article-title":"Indian hedgehog synchronizes skeletal angiogenesis and perichondrial maturation with cartilage development","volume":"132","author":"Colnot","year":"2005","journal-title":"Development"},{"issue":"19","key":"2024020319001019700_jbmr2582-bib-0015","doi-asserted-by":"crossref","first-page":"4339","DOI":"10.1242\/dev.02025","article-title":"Ihh controls cartilage development by antagonizing Gli3, but requires additional effectors to regulate osteoblast and vascular development","volume":"132","author":"Hilton","year":"2005","journal-title":"Development"},{"issue":"1","key":"2024020319001019700_jbmr2582-bib-0016","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1242\/dev.01564","article-title":"Sequential roles of Hedgehog and Wnt signaling in osteoblast development","volume":"132","author":"Hu","year":"2005","journal-title":"Development"},{"issue":"18","key":"2024020319001019700_jbmr2582-bib-0017","doi-asserted-by":"crossref","first-page":"3695","DOI":"10.1242\/dev.02546","article-title":"Wnt\/beta-catenin signaling interacts differentially with Ihh signaling in controlling endochondral bone and synovial joint formation","volume":"133","author":"Mak","year":"2006","journal-title":"Development"},{"issue":"16","key":"2024020319001019700_jbmr2582-bib-0018","doi-asserted-by":"crossref","first-page":"3231","DOI":"10.1242\/dev.02480","article-title":"Distinct roles for Hedgehog and canonical Wnt signaling in specification, differentiation and maintenance of osteoblast progenitors","volume":"133","author":"Rodda","year":"2006","journal-title":"Development"},{"issue":"33","key":"2024020319001019700_jbmr2582-bib-0019","doi-asserted-by":"crossref","first-page":"12097","DOI":"10.1073\/pnas.1302703111","article-title":"Hypertrophic chondrocytes can become osteoblasts and osteocytes in endochondral bone formation","volume":"111","author":"Yang","year":"2014","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"19","key":"2024020319001019700_jbmr2582-bib-0020","doi-asserted-by":"crossref","first-page":"1959","DOI":"10.1056\/NEJMoa044264","article-title":"Circulating osteoblast-lineage cells in humans","volume":"352","author":"Eghbali-Fatourechi","year":"2005","journal-title":"N Engl J Med"},{"issue":"5","key":"2024020319001019700_jbmr2582-bib-0021","doi-asserted-by":"crossref","first-page":"1370","DOI":"10.1016\/j.bone.2006.12.064","article-title":"Characterization of circulating osteoblast lineage cells in humans","volume":"40","author":"Eghbali-Fatourechi","year":"2007","journal-title":"Bone"},{"issue":"3","key":"2024020319001019700_jbmr2582-bib-0022","doi-asserted-by":"crossref","first-page":"e1001505","DOI":"10.1371\/journal.pbio.1001505","article-title":"Secretion and signaling activities of lipoprotein-associated hedgehog and non-sterol-modified hedgehog in flies and mammals","volume":"11","author":"Palm","year":"2013","journal-title":"PLoS Biol"},{"issue":"23","key":"2024020319001019700_jbmr2582-bib-0023","doi-asserted-by":"crossref","first-page":"2636","DOI":"10.1101\/gad.249763.114","article-title":"Production of systemically circulating Hedgehog by the intestine couples nutrition to growth and development","volume":"28","author":"Rodenfels","year":"2014","journal-title":"Genes Dev"},{"issue":"3","key":"2024020319001019700_jbmr2582-bib-0024","first-page":"477","article-title":"urakami S","volume":"386","author":"Kawanami","year":"2009","journal-title":"Mice expressing GFP and CreER in osteochondro progenitor cells in the periosteum. 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