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Ono -- PTH-Pthrp Receptor Signaling in Osterix-Expressing Progenitors.Pdf 3/27/14 Root forma)on Cementum Dentin Cementoblast Odontoblast Role of PTH/PTHrP Receptor Signaling on Root Epithelial rests Formaon of Malassez (ERM) Dental Wanida Ono papilla cells AGE Orthodon;cs, Department of Developmental Biology, Harvard School of Dental Medicine Endocrine Unit, MassachuseLs General Hospital and Harvard Medical School Dental follicle cells Hertwig’s epithelial root sheath (HERS) Osterix PTHrP-PPR system • Transcripon factor essen;al to osteoblast differen;aon PTHrP (Nakashima K et al. 2002) PTH/PTHrP receptor (PPR) • Expressed in odontoblasts and alveolar osteoblasts during Gαs Gq tooth development (Chen S et al. 2009) • Controls cellular cementum formaon (Cao Z et al. 2012) • Mediates epithelial-mesenchymal interacons • Osterix-expressing precursors in the perichondrium move • PTHrP is expressed in enamel epithelia/HERS? to bone marrow and become osteoblasts during fetal • PPR is expressed in dental mesenchymes development (Maes C, Kronenberg HM et al. 2010) (Beck et al 1995; Lee Deeds and Segre 1995; Liu et al 1998) • PTHrP is required for tooth erup;on in mice (Philbrick WM, Karaplis AC et al. PNAS 1998) Osterix+ Root-forming • PPR haploinsufficiency is associated with primary cells progenitors failure of tooth erup;on (PFE) in humans ? (Decker E, Weber BH et al. Am J Hum Gen 2008) PTHrP expression paern during root morphogenesis Aims of this study PTHrPLacZ/+ x40 P7 P14 P49 • Iden;fy how osterix-expressing progenitors contribute to murine root morphogenesis • Understand how PTH/PTHrP receptor signal regulates root-forming progenitors PTHrP-LacZ x200 P7 x400 P14 ? PTHrP Epithelial root sheath PPR PTHrP-LacZ Osx+ progenitors Odontoblast PTHrP par;cipates in …….. Dental follicle/papilla cells Cementoblast - dental (root) development - homeostasis of root-bone interface ( composed by PDL, cementum) 1 3/27/14 Experimental approach (1) Working hypothesis • Iden;fy how osterix-expressing progenitors contribute to murine root formaon PTHrP HERS (Epithelial root sheath) Tamoxifen Osx-CreERT2 PPR ? Molar root forma+on P3 P4 P7 P11 P25 Root-forming progenitors Cementoblast CAG Tomato Dental follicle cells X loxP Stop loxP Cre CAG Tomato Rosa26-Tomato In vivo fate mapping of Osx+ cells Osx+ cells are root-forming progenitors Tam P3 P4 P7 P14 P25 Osx+ cells Dental pulp cells Osx-CreER; Tomato P4 (+1d) Dental papilla Odontoblasts Osx+ cells Periodontal ligament cells Dental follicle Postnatal day 4 Cementoblasts Mandibular M1, mid-sagiLal sec;on Alveolar osteoblasts Experimental approach (2) Osx-Cre; PPRflox/+ PPRflox/flox • Understand how PTH/PTHrP receptor signal (PPR) X regulates root-forming progenitors P7 Control CondiIonal Het Condional KO Osx-Cre; PPRflox/+ PTH/PTHrP receptor (PPR) locus E1 E2 PPR floxed allele X Cre (Kobayashi T, Kronenberg HM et al. 2002) E2 PPR null allele PPRflox/flox Osx-Cre; PPRflox/+ Osx-Cre; PPRflox/flox PPRflox/flox 2 3/27/14 PPR signal in Osx+ progenitors are Conclusion essenal for root formaon • Osterix-expressing cells are dental root-forming progenitors during morphogenesis: Osx+ cells PPR signal Osx+ cells become dental pulp cells and odontoblasts, Periodontal ligament cells periodontal ligament cells, cementoblasts and alveolar osteoblasts Dental follicle • Deleon of PTH/PTHrP receptor in Osx+ cells leads to Cementoblasts dental root malformaon and failure of erupon 1) Truncaon of the dental root PPR signal 2) Hypogenesis of periodontal ligament 3) Ankylosis of cementum to bone 4) Underdeveloped mandible 5) Failure of molar erup;on “malformed/ truncated root” Acknowledgement Endocrine Unit / MassachuseLs General Hospital Hank Kronenberg Noriaki Ono Shigeki Nishimori Tatsuya Kobayashi Marc Wein Forest Lai Everyone in the Endocrine Unit HSDM AGE Orthodon;cs Program Acknowledgement for Important Mice… Arthur Broadus PTHrP-LacZ Andrew McMahon Osx-Cre::GFP 3 .
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