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Root forma on
Cementum Dentin Cementoblast Odontoblast
Role of PTH/PTHrP Receptor Signaling on Root Epithelial rests Forma on of Malassez (ERM) Dental Wanida Ono papilla cells
AGE Orthodon cs, Department of Developmental Biology, Harvard School of Dental Medicine
Endocrine Unit, Massachuse s General Hospital and Harvard Medical School Dental follicle cells
Hertwig’s epithelial root sheath (HERS)
Osterix PTHrP-PPR system • Transcrip on factor essen al to osteoblast differen a on 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 forma on (Cao Z et al. 2012) • Mediates epithelial-mesenchymal interac ons • 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 pa ern 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)
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Experimental approach (1) Working hypothesis • Iden fy how osterix-expressing progenitors contribute to murine root forma on 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-sagi al 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 Condi onal Het Condi onal 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
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PPR signal in Osx+ progenitors are Conclusion
essen al for root forma on • 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 • Dele on of PTH/PTHrP receptor in Osx+ cells leads to
Cementoblasts dental root malforma on and failure of erup on 1) Trunca on 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 / Massachuse s 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
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