Role of GSK-3β in the Osteogenic Differentiation of Palatal Mesenchyme 英文参考文献.docVIP

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Role of GSK-3β in the Osteogenic Differentiation of Palatal Mesenchyme 英文参考文献.doc

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Role of GSK-3β in the Osteogenic Differentiation of Palatal Mesenchyme 英文参考文献

RoleofGSK-3bintheOsteogenicDifferentiationof PalatalMesenchyme EmilyR.Nelson1.,BenjaminLevi1.,MichaelSorkin1,AaronW.James1,KarenJ.Liu2,NatalinaQuarto1,3, MichaelT.Longaker1,4 * 1Hagey Laboratory for Pediatric Regenerative Medicine, Plastic and Reconstructive Surgery Division, Department of Surgery, Stanford University School of Medicine, Stanford,California,UnitedStatesofAmerica,2DepartmentofCraniofacialDevelopment,King’sCollegeLondon,London,UnitedKingdom,3DipartimentodiScienze Chirurgiche, Anestesiologiche-Rianimatorie e dell ‘Emergenza ‘‘Giuseppe Zannini,’’ Universita’ degli Studi di Napoli Federico II, Napoli, Italy, 4Institute for Stem Cell BiologyandRegenerativeMedicine,StanfordUniversity,Stanford,California,UnitedStatesofAmerica Abstract Introduction:ThefunctionofGlycogenSynthaseKinases3b(GSK-3b)haspreviouslybeenshowntobenecessaryfornormal secondarypalatedevelopment.UsingGSK-3?nullmouseembryos,weexaminethepotentialcoordinaterolesofWntand Hedgehogsignalingonpalatalossification. Methods:PalateswereharvestedfromGSK-3b,embryonicdays15.0–18.5(e15.0–e18.5),ande15.5IndianHedgehog(Ihh) null embryos, and their wild-type littermates. The phenotype of GSK-3b null embryos was analyzed with skeletal whole mountandpentachromestains.Spatiotemporalregulationofosteogenicgeneexpression,inadditiontoWntandHedgehog signalingactivity,wereexaminedinvivoonGSK-3bandIhh+/+and2/2e15.5embryosusinginsituhybridizationand immunohistochemistry.Tocorroboratetheseresults,expressionofthesamemoleculartargetswereassessedbyqRT-PCRof e15.5palates,ore13.5palateculturestreatedwithbothWntandHedgehogagonistsandanatagonists. Results:GSK-3bnullembryosdisplayeda48percentdecrease(*p,0.05)inpalatineboneformationcomparedtowild-type littermates.GSK-3bnullembryosalsoexhibiteddecreasedosteogenicgeneexpressionthatwasassociatedwithincreased WntanddecreasedHedgehogsignaling.e13.5palateculturestudiesdemonstratedthatWntsignalingnegativelyregulates both osteogenic gene expression and Hedgehog signaling activity, wh