小鼠早期胚胎发育专业材料.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1专业知识体轴形成和原肠运动体轴形成和原肠运动Axis formation & Gastrulation2专业知识小鼠早期胚胎发育小鼠早期胚胎发育3专业知识Early mouse developmentPreimplantation4专业知识PostimplantationEarly mouse development(early) Epiblast(late) EpiblastAnterior neuroectodermVE: visceral endodermAVE: anterior VEDVE: distal VE5专业知识Cell lineages in early mouse embryoMorulaInner Cell MassTrophectodermPrimitive endodermEpiblastParietal endodermVisceral endodermDefinitive endodermMesodermEctodermMesendodermCNSEpidermis6专业知识Mouse embryo development from fertilization to gastrulationDevelopment 139, 3-14, 2012？？？？？？7专业知识Two possible mechanisms for the generation of multiple cell types and polarityDevelopment 139, 3-14, 20128专业知识Dev Cell 16, 398-410, 2009Mouse trophectoderm distinguish from inner cell mass regulated by Hippo signaling pathway 9专业知识Cell position-dependent differentiation mediated by Hippo signalingDevelopment 139, 3-14, 201210专业知识Mouse embryo development from fertilization to gastrulationDevelopment 139, 3-14, 2012？？11专业知识Early mouse developmentE3.5E4.5ICM12专业知识‘‘Salt and Pepper’’ expression of Nanog and Gata6 in the ICM of E3.5 blastocystsDev Cell 10, 615-624, 200613专业知识Segregation and relocation of epiblast and PrE cells Development 139, 3-14, 201214专业知识Mouse embryo development from fertilization to gastrulationDevelopment 139, 3-14, 2012？？15专业知识Emerging asymmetry and establishment of anterior-posterior polarity in the E5.75 mouse embryoAxis of Anterior-PosteriorAxis of Proximal & Distal16专业知识Distal VE to AVECell movements in early mouse embryosEpiblast to posteriorExtraembryonic ectoderm to posterior epiblastEpiblast to posterior17专业知识Otx2 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 endoderm18专业知识Asymmetric expression of Lefty1 transgenes in peri-implantation mouse embryosDev Cell 10, 451-459, 200619专业知识Two types of Lefty1+ cell in mouse embryos between E3.5 and E4.5Nat Cell Biol 13, 743-752, 201120专业知识Distinct origins of DVE and AVENat Cell Biol 13, 743-752, 201121专业知识Origins of the dorsal and anterior visceral endodermDevelopment 139, 3-14, 201222专业知识Signaling pathways involved in A-P axis determinationDevelopment 139, 3-14, 201223专业知识Mouse embryo development from fertilization to gastrulationDevelopment 139, 3-14, 201224专业知识Nervous systemSkin and hairHeart Muscle liver Lungs … … … … Early mouse embryo development and lineage segregation3.5dpcICM 4.5dpcearly epiblast5.5dpclate epiblast6.5dpcPrimitive streak25专业知识Rapid 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, 198626专业知识VE: visceral endodermAVE: anterior VEDVE: distal VEPrimitive streak formation PS: Primitive Streak27专业知识EMT at mouse primitive streak28专业知识EMT at mouse primitive streakEpiblastPrimitve StreakNascent MesodermCell Adhesion & Migration 4:3, 447-45729专业知识Mouse gastrulation and germ layer formationCell 132, 661–680, 200830专业知识MyocardinE7.5E8.5Late mouse developmentAxis of Dorsal-Ventral31专业知识Cell lineages in early mouse embryo32专业知识Embryo culture system for time-lapse movie acquisition of node morphogenesisDev Cell 13, 884-896, 2007in vitro embryo cultureGFP knock-in tracing 33专业知识Time-lapse images of node morphogenesisNoto-Dev Cell 13, 884-896, 200734专业知识The displacement model of endoderm morphogenesis in the mouse gastrula35专业知识The dispersal model of endoderm morphogenesis in the mouse gastrula36专业知识Visceral endoderm-derived cells are incorporated into the gut tube12–18 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, 200837专业知识Cell lineages in early mouse embryo38专业知识Definitive endodermMesodermEctodermMesendodermCNSEpidermisEpiblastCell lineage in early mouse embryoE7.0TEctodermMesendoderm39专业知识BraWnt3E7.5Mesendoderm markers 40专业知识Gene expression and regionalized allocation of tissue progenitorsAxial mesodermDefinitive endodermParaxial mesodermLateral plate mesodermExtraembryonic mesodermNeuroectoderm?41专业知识The markers of germ layersDefinitive endodermMesodermEctodermMesendodermNeuralEpidermisNkx2.5Sox17Nodal, T, Wnt3, GscEpiblastMesp1Mixl1Flk1Gata4Gata6Foxa2Sox1Pax6NestinSix3Ck5Ck8Ck18Zo-1FGF5, Nodal42专业知识Fate-mapping of the distal cap epiblast by carbocyanine dye labelingE6.5Dye injection43专业知识The distal cap epiblast cells migrate to the anterior neuroectoderm44专业知识 Development, 107, 55-67 (1989)Current Biology 1996, Vol 6 No 2Fate maps for the ectodermal layer Wheat germ agglutinin (WGA)45专业知识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 hr46专业知识Development 139, 423-436 (2012)Tissue contributions of descendant cells according to the original position of HRP-injected progenitors47专业知识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48专业知识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 49专业知识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 2010IdpErk50专业知识In vitroESCs ESD-EpiSCs Ectodermal cellNPCEpidermal cellMesendodermal cellEndoderm cellMesoderm cell Ectodermal and mesendodermal stem cells？？51专业知识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 tissue52专业知识The 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53专业知识The anterior epiblast in E7.0 mouse embryo has the potential to give rise to neural and epidermal cellsABCD54专业知识Regionalization of anterior epiblast in E7.0 mouse embryo55专业知识Ectoderm formation during mouse embryonic development and stem cell differentiationLi et al. Development 201356专业知识Thanks!57专业知识。