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3/27/14

Root forma)on

Cementum Dentin

  • Cementoblast
  • Odontoblast

Role of PTH/PTHrP Receptor Signaling on Root
Formaꢀon

Epithelial rests of Malassez

(ERM)

Dental papilla cells

Wanida Ono

AGE Orthodonꢀcs, 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

PTHrP

•ꢀ Transcrip)on factor essenꢀal to osteoblast differenꢀaꢀon

(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 formaꢀon

(Cao Z et al. 2012)

•ꢀ Mediates epithelial-mesenchymal interac)ons

•ꢀ Osterix-expressing precursors in the perichondrium move

to bone marrow and become osteoblasts during fetal

•ꢀ PTHrP is expressed in enamel epithelia/HERS? •ꢀ PPR is expressed in dental mesenchymes

(Beck et al 1995; Lee Deeds and Segre 1995; Liu et al 1998)

development

(Maes C, Kronenberg HM et al. 2010)

•ꢀ PTHrP is required for tooth erupꢀon in mice

(Philbrick WM, Karaplis AC et al. PNAS 1998)

Osterix+ cells

Root-forming progenitors

•ꢀ PPR haploinsufficiency is associated with primary

?

failure of tooth erupꢀon (PFE) in humans

(Decker E, Weber BH et al. Am J Hum Gen 2008)

PTHrP expression paꢀern during root morphogenesis

Aims of this study

•ꢀ Idenꢀfy how osterix-expressing progenitors contribute

PTHrPLacZ/+

x40

  • P7
  • P14
  • P49

to murine root morphogenesis
•ꢀ Understand how PTH/PTHrP receptor signal regulates root-forming progenitors

PTHrP--acZ

  • x200
  • x400

  • P7
  • P14

PTHrP

Epithelial root sheath

?

PPR

PTHrP--acZ

Osx+ progenitors

Odontoblast Cementoblast

PTHrP parꢀcipates in ……..
-ꢀdental (root) development -ꢀhomeostasis of root-bone interface (composed by PDL, cementum)

Dental follicle/papilla cells

1
3/27/14

Experimental approach (1)

Working hypothesis

•ꢀ Idenꢀfy how osterix-expressing progenitors contribute

to murine root forma)on

PTHrP

HERS (Epithelial root sheath)

Tamoxifen

Osx-CreERT2

?

Molar root forma+on

PPR

P3 P4 P7

  • P11
  • P25

Root-forming progenitors

CAG

Cementoblast

Tomato

loxP Stop loxP

X

Dental follicle cells

Cre

CAG

Tomato

Rosa26-Tomato

Osx+ cells are root-forming progenitors

In vivo fate mapping of Osx+ cells

Tam

  • P3 P4
  • P7

Osx+ cells

  • P14
  • P25

Dental pulp cells

Osx-CreER; Tomato

P4 (+1d)

Dental papilla

Odontoblasts

Osx+ cells

Periodontal ligament cells

Dental follicle

Postnatal day 4 Mandibular M1, mid-sagiLal secꢀon

Cementoblasts Alveolar osteoblasts

Experimental approach (2)

  • Osx-Cre ; PPRflox /+
  • PPRflox/flox

X

•ꢀ Understand how PTH/PTHrP receptor signal (PPR)

regulates root-forming progenitors

P7

  • Control
  • CondiIonal Het

CondiIonal KO

Osx-Cre ; PPRflox/+

PTH/PTHrP receptor (PPR) locus

  • E1
  • E2

PPR floxed allele

Cre

(Kobayashi T, Kronenberg HM et al. 2002)

X

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 essen)al for root forma)on

Conclusion

•ꢀ Osterix-expressing cells are dental root-forming progenitors during morphogenesis:

Osx+ cells become dental pulp cells and odontoblasts,

Osx+ cells

PPR signal

Periodontal ligament cells

periodontal ligament cells, cementoblasts and alveolar osteoblasts

•ꢀ Deleꢀon of PTH/PTHrP receptor in Osx+ cells leads to

Dental follicle

dental root malforma)on and failure of erup)on

Cementoblasts

1)ꢀ Truncaꢀon of the dental root 2)ꢀ Hypogenesis of periodontal ligament 3)ꢀ Ankylosis of cementum to bone 4)ꢀ Underdeveloped mandible

PPR signal

5)ꢀ Failure of molar erupꢀon

“malformed/ truncated root”

Acknowledgement

Endocrine Unit / MassachuseLs General Hospitalꢀ

Hank Kronenberg Noriaki Ono Shigeki Nishimori Tatsuya Kobayashi Marc Wein Forest Lai Everyone in the Endocrine Unit

HSDM AGE Orthodonꢀcs Program Acknowledgement for Important Mice…ꢀ

Arthur Broadus
PTHrP-LacZ
Andrew McMahon
Osx-Cre::GFP

3

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