小鼠早期胚胎发育.ppt

小鼠早期胚胎发育小鼠早期胚胎发育景乃禾景乃禾中国科学院上海生物化学与细胞生物学研究所中国科学院上海生物化学与细胞生物学研究所体轴形成和原肠运动体轴形成和原肠运动Axis formation&Gastrulation小鼠早期胚胎发育小鼠早期胚胎发育Early mouse developmentPreimplantationPostimplantationEarly mouse development(early)Epiblast(late)EpiblastAnterior neuroectodermVE:visceral endodermAVE:anterior VEDVE:distal VECell lineages in early mouse embryoMorulaInner Cell MassTrophectodermPrimitive endodermEpiblastParietal endodermVisceral endodermDefinitive endodermMesodermEctodermMesendodermCNSEpidermisMouse embryo development from fertilization to gastrulationDevelopment 139,3-14,2012？Two possible mechanisms for the generation of multiple cell types and polarityDevelopment 139,3-14,2012Dev Cell 16,398-410,2009Mouse trophectoderm distinguish from inner cell mass regulated by Hippo signaling pathway Cell position-dependent differentiation mediated by Hippo signalingDevelopment 139,3-14,2012Mouse embryo development from fertilization to gastrulationDevelopment 139,3-14,2012？Early mouse developmentE3.5E4.5ICMSalt and Pepper expression of Nanog and Gata6 in the ICM of E3.5 blastocystsDev Cell 10,615-624,2006Segregation and relocation of epiblast and PrE cells Development 139,3-14,2012Mouse embryo development from fertilization to gastrulationDevelopment 139,3-14,2012？Emerging asymmetry and establishment of anterior-posterior polarity in the E5.75 mouse embryoAxis of Anterior-PosteriorAxis of Proximal&DistalDistal VE to AVECell movements in early mouse embryosEpiblast to posteriorExtraembryonic ectoderm to posterior epiblastEpiblast to posteriorOtx2 VEcis-lacZ5.5 dpc5.75 dpc6.25 dpc6.0 dpcOtx2 KI-lacZMovement of Otx2-positive cells from DVE to AVEDVE:distal visceral endoderm;AVE:anterior visceral endodermAsymmetric expression of Lefty1 transgenes in peri-implantation mouse embryosDev Cell 10,451-459,2006Two types of Lefty1+cell in mouse embryos between E3.5 and E4.5Nat Cell Biol 13,743-752,2011Distinct origins of DVE and AVENat Cell Biol 13,743-752,2011Origins of the dorsal and anterior visceral endodermDevelopment 139,3-14,2012Signaling pathways involved in A-P axis determinationDevelopment 139,3-14,2012Mouse embryo development from fertilization to gastrulationDevelopment 139,3-14,2012Nervous systemSkin and hairHeart Muscle liver Lungs Early mouse embryo development and lineage segregation3.5dpcICM 4.5dpcearly epiblast5.5dpclate epiblast6.5dpcPrimitive streakRapid proliferation during gastrulation:dramatic signaling and gene expression changes Fossat,N.,et al.(2012)Cell Cycle 11:26-32“It is not birth,marriage,or death,but gastrulation,which is truly the most important time in your life.”Lewis Wolpert,1986VE:visceral endodermAVE:anterior VEDVE:distal VEPrimitive streak formation PS:Primitive StreakEMT at mouse primitive streakEMT at mouse primitive streakEpiblastPrimitve StreakNascent MesodermCell Adhesion&Migration 4:3,447-457Mouse gastrulation and germ layer formationCell 132,661680,2008MyocardinE7.5E7.5E8.5E8.5Late mouse developmentAxis of Dorsal-VentralCell lineages in early mouse embryoEmbryo culture system for time-lapse movie acquisition of node morphogenesisDev Cell 13,884-896,2007in vitro embryo cultureGFP knock-in tracing Time-lapse images of node morphogenesisNoto-Dev Cell 13,884-896,2007The displacement model of endoderm morphogenesis in the mouse gastrulaThe dispersal model of endoderm morphogenesis in the mouse gastrulaVisceral endoderm-derived cells are incorporated into the gut tube1218 somite embryos revealed the presence of Afp-GFP+cells in the gut tubeThis was not new expression of the transgene,as GFP mRNA could only be detectedin the yolk sac at this stageThese visceral endoderm derived cells continue to divide and are eventually incorporatedinto the epithelium of the gut tubeDev Cell 15,509-520,2008Cell lineages in early mouse embryoDefinitive endodermMesodermEctodermMesendodermCNSEpidermisEpiblastCell lineage in early mouse embryoE7.0TEctodermMesendodermBraWnt3E7.5Mesendoderm markers Gene expression and regionalized allocation of tissue progenitorsAxial mesodermDefinitive endodermParaxial mesodermLateral plate mesodermExtraembryonic mesodermNeuroectoderm?The markers of germ layersDefinitive endodermMesodermEctodermMesendodermNeuralEpidermisNkx2.5Sox17Nodal,T,Wnt3,GscEpiblastMesp1Mixl1Flk1Gata4Gata6Foxa2Sox1Pax6NestinSix3Ck5Ck8Ck18Zo-1FGF5,NodalFate-mapping of the distal cap epiblast by carbocyanine dye labelingE6.5Dye injectionThe distal cap epiblast cells migrate to the anterior neuroectoderm Development,107,55-67(1989)Current Biology 1996,Vol 6 No 2Fate maps for the ectodermal layer Wheat germ agglutinin(WGA)Development 139,423-436(2012)Single cell fate map of ectodermal cells in early mouse embryoE7.0,single cell injection of HRP and LRDX,and in vitro culture 24 hrDevelopment 139,423-436(2012)Tissue contributions of descendant cells according to the original position of HRP-injected progenitors内胚层内胚层中胚层中胚层外胚层外胚层中内胚层中内胚层神经外胚层神经外胚层表面外胚层表面外胚层上胚层上胚层囊胚（内细胞团）囊胚（内细胞团）受精卵受精卵多能干细胞的细胞谱系多能干细胞的细胞谱系In vivoIn vitroICM/epiblast(E3.5-4.5)Late epiblast (E5.5)(E6.5)Three germ layersEctoderm Endoderm Mesoderm ESCs ESD-EpiSCs Ectodermal cellNPCEpidermal cellMesendodermal cell多能性细胞谱系的建立Endoderm cellMesoderm cell ICM/epiblast(E3.5-4.5)Blastocyte stageEgg cylinder stageEpiblast(E5.5)Neural plate(E7.5)GastrulationIn vivoIn vitroNPCsESCsESD-EpiSCsBMPFGF4pErkNon-neural lineage cellsBMPIdBMP signaling functions distinctly at the two stages during neural differentiation of ESCsZhang and Li et.al.Development 2010IdpErkIn vitroESCs ESD-EpiSCs Ectodermal cellNPCEpidermal cellMesendodermal cellEndoderm cellMesoderm cell Ectodermal and mesendodermal stem cells？PSE6.0ES-MS E6.5OB-EB E7.0 LB E7.5EHF-LHFE7.75-8.0Marker genes expression during early mouse embryonic development Hesx1Fgf5Six3Oct6Sox2TE7.0E7.5HypothesisEctoderm Cells:E7.0 Anterior part of epiblast tissueThe anterior part of epiblasts at E6.5 and E7.5 does not have ectoderm differentiation potential外胚层外胚层神经外胚层神经外胚层表面外胚层表面外胚层The anterior epiblast in E7.0 mouse embryo has the potential to give rise to neural and epidermal cellsABCDRegionalization of anterior epiblast in E7.0 mouse embryoEctoderm formation during mouse embryonic development and stem cell differentiationLi et al.Development 2013Thanks!