小鼠早期胚胎发育课件.ppt

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1、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 VEC

2、ell 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 t

3、he 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 develop

4、ment 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 developmen

5、t 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 ectoder

6、m 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-

7、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 determinationDevelopmen

8、t 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 duri

9、ng 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 formatio

10、n 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 i

11、n 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 gastru

12、laThe 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

13、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 embryoE

14、7.0TEctodermMesendodermBraWnt3E7.5Mesendoderm markers Gene expression and regionalized allocation of tissue progenitorsAxial mesodermDefinitive endodermParaxial mesodermLateral plate mesodermExtraembryonic mesodermNeuroectoderm?The markers of germ layersDefinitive endodermMesodermEctodermMesendoderm

15、NeuralEpidermisNkx2.5Sox17Nodal,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

16、)M agazi ne105Fi gure 1Fat e maps f or embryos of f our vert ebratespeci es,drawn w i t h t hei r normal t opography(see t ext f or f urt her det ai l s).(a)Appearance ofeach embryo bef ore gast rul at i on.Scal e bars:f i sh,500 600 mm;f rog 1 2m m;chi ck,2-3m m;mouse,700 mm.(b)Fat e maps at earl y

17、gast rul at i on,draw n i n normal t opography.Embryos are al i gned so t hat t he organi zer i son t he ri ght hand si de of each f at e map.Theorgani zer i s f ound i n t he embryoni c shi el d(SH)of t he zebraf i sh and t he dorsalbl ast opore l i p (BL)of t he f rog embr yo;t heprecursor of t he

18、 organi zer(t he node)i s f i rstf ound as an i nt egral part of t he earl y primi t i vest reak(PS),t he f i rst morphol ogi cal l andmarkof t he chi ck or mouse embryo.I n represent i ngt he chi ck embr yo,separate images of a bl ast o-derm are show n t o i ndi cate germ l ayerprecursors w i t h o

19、verl appi ng di st ri but i ons.(c)Fat e maps f or t he ect odermal l ayers at t he endof gast rul at i on(mesodermal and endodermall ayers are not shown due t o spaceconst rai nt s).The ant eri or post eri or axes areal i gned hori zont al l y i n t he f i gure,such t hat t het he si t e of cel l m

20、ovement t o t he i nt eri or i s ont he post eri or(ri ght hand si de)of t he map.Byt hi s stage,t he organi zer i s morphol ogi cal l ydi st i ngui shabl e at t he ant eri or ext remi t y of t heprimi t i ve st reak(N),as t he H ensen s node(chi ck)or t he node(mouse).Ect odermalt i ssues:ne,neuroe

21、ct oderm,f b,f orebrai n;mb,mi dbrai n;hb,hi ndbrai n;sc,spi nal cord;se,surf ace ect oderm.M esodermalt i ssues:pxm,paraxi almesoderm;hm,head mesoderm;sm,somi tes;lm,l at eral mesoderm;nd,not ochord;ht,heart;bl,bl ood.Endodermal t i ssues:end,gut endoderm.Ext raembr yoni c t i ssues:am,amni on ect

22、oderm;exm,ext raembryoni cmesoderm.Fi shFrogChi ckM ouseAnimal pol eAnimal pol eProximalVegetal pol eVegetal pol eDi st alBl ast ocoelBl ast odermBl ast odermEpi bl astYol k cel l syol k(a)(c)(b)50%epi bol y8000 cel l s i n bl ast oderm90%epi bol yStage 10+5000 cel l s i n bl ast ul aSt age 12.5 13S

23、t age XI V600 700 cel l s i n epi bl astSt age 56.5 days post coi t um650 cel l s i n epi bl ast7.5 days post coi t umYol kEarl y embr yo Ect oderm M esoderm Endoderm Organi zer Yol k Ext raembr yoni c t i ssues 1996 Cur rent Bi ol ogysesepxmlmhtexmamnendPSPSendendndneneneyol ksesesepxmpxmhthtlmblbl

24、ndBLlmlmsmhmhtndSHexmendf bf bmbsehbscscNPSexmlmpxmscscsesehbhbmbmbf bf bmbhbpxmndNlmPSexmendM agazi ne105Fi gure 1Fate maps f or embryos of f our vertebratespeci es,drawn w i t h t hei r normal t opography(see text f or f urt her detai l s).(a)Appearance ofeach embryo bef ore gast rul at i on.Scal

25、e bars:f i sh,500 600 mm;f rog 1 2m m;chi ck,2-3m m;mouse,700 mm.(b)Fate maps at earl ygast rul at i on,draw n i n normal t opography.Embryos are al i gned so t hat t he organi zer i son t he ri ght hand si de of each f ate map.Theorgani zer i s f ound i n t he embryoni c shi el d(SH)of t he zebraf

26、i sh and t he dorsalbl ast opore l i p (BL)of t he f rog embryo;t heprecursor of t he organi zer(t he node)i s f i rstf ound as an i ntegral part of t he earl y primi t i vest reak(PS),t he f i rst morphol ogi cal l andmarkof t he chi ck or mouse embryo.I n represent i ngt he chi ck embryo,separate

27、images of a bl ast o-derm are show n t o i ndi cate germ l ayerprecursors w i t h overl appi ng di st ri but i ons.(c)Fate maps f or t he ect odermal l ayers at t he endof gast rul at i on(mesodermal and endodermall ayers are not shown due t o spaceconst rai nt s).The anteri or posteri or axes areal

28、 i gned hori zontal l y i n t he f i gure,such t hat t het he si te of cel l movement t o t he i nteri or i s ont he posteri or(ri ght hand si de)of t he map.Byt hi s stage,t he organi zer i s morphol ogi cal l ydi st i ngui shabl e at t he anteri or ext remi t y of t heprimi t i ve st reak(N),as t

29、he H ensen s node(chi ck)or t he node(mouse).Ect odermalt i ssues:ne,neuroect oderm,f b,f orebrai n;mb,mi dbrai n;hb,hi ndbrai n;sc,spi nal cord;se,surf ace ect oderm.M esodermalt i ssues:pxm,paraxi almesoderm;hm,head mesoderm;sm,somi tes;lm,l ateral mesoderm;nd,not ochord;ht,heart;bl,bl ood.Endoder

30、mal t i ssues:end,gut endoderm.Ext raembryoni c t i ssues:am,amni on ect oderm;exm,ext raembryoni cmesoderm.Fi shFrogChi ckM ouseAnimal pol eAnimal pol eProximalVegetal pol eVegetal pol eDi stalBl ast ocoelBl ast odermBl ast odermEpi bl astYol k cel l syol k(a)(c)(b)50%epi bol y8000 cel l s i n bl a

31、st oderm90%epi bol yStage 10+5000 cel l s i n bl ast ul aSt age 12.5 13St age XI V600 700 cel l s i n epi bl astSt age 56.5 days post coi t um650 cel l s i n epi bl ast7.5 days post coi t umYol kEarl y embr yo Ect oderm M esoderm Endoderm Organi zer Yol k Ext raembr yoni c t i ssues 1996 Cur rent Bi

32、 ol ogysesepxmlmhtexmamnendPSPSendendndneneneyol ksesesepxmpxmhthtlmblblndBLlmlmsmhmhtndSHexmendf bf bmbsehbscscNPSexmlmpxmscscsesehbhbmbmbf bf bmbhbpxmndNlmPSexmendM agazine105Fi gure 1Fate maps f or embryos of f our vertebratespeci es,drawn w i t h t hei r normal t opography(see text f or f urt he

33、r detai l s).(a)Appearance ofeach embryo bef ore gast rul at i on.Scal e bars:f i sh,500 600 mm;f rog 1 2m m;chi ck,2-3m m;mouse,700 mm.(b)Fate maps at earl ygast rul at i on,draw n i n normal t opography.Embryos are al i gned so t hat t he organi zer i son t he ri ght hand si de of each f ate map.T

34、heorgani zer i s f ound i n t he embryoni c shi el d(SH)of t he zebraf i sh and t he dorsalbl ast opore l i p (BL)of t he f rog embryo;t heprecursor of t he organi zer(t he node)i s f i rstf ound as an i ntegral part of t he earl y primi t i vest reak(PS),t he f i rst morphol ogi cal l andmarkof t h

35、e chi ck or mouse embryo.I n represent i ngt he chi ck embryo,separate images of a bl ast o-derm are show n t o i ndi cate germ l ayerprecursors w i t h overl appi ng di st ri but i ons.(c)Fate maps f or t he ect odermal l ayers at t he endof gast rul at i on(mesodermal and endodermall ayers are not

36、 show n due t o spaceconst rai nt s).The anteri or posteri or axes areal i gned hori zontal l y i n t he f i gure,such t hat t het he si te of cel l movement t o t he i nteri or i s ont he posteri or(ri ght hand si de)of t he map.Byt hi s stage,t he organi zer i s morphol ogi cal l ydi st i ngui sha

37、bl e at t he anteri or ext remi t y of t heprimi t i ve st reak(N),as t he H ensen s node(chi ck)or t he node(mouse).Ect odermalt i ssues:ne,neuroect oderm,f b,f orebrai n;mb,mi dbrai n;hb,hi ndbrai n;sc,spi nal cord;se,surf ace ect oderm.M esodermalt i ssues:pxm,paraxi almesoderm;hm,head mesoderm;s

38、m,somi tes;lm,l ateral mesoderm;nd,not ochord;ht,heart;bl,bl ood.Endodermal t i ssues:end,gut endoderm.Ext raembryoni c t i ssues:am,amni on ect oderm;exm,ext raembryoni cmesoderm.Fi shFrogChi ckM ouseAnimal pol eAnimal pol eProximalVegetal pol eVegetal pol eDi stalBl ast ocoelBl ast odermBl ast ode

39、rmEpi bl astYol k cel l syol k(a)(c)(b)50%epi bol y8000 cel l s i n bl ast oderm90%epi bol yStage 10+5000 cel l s i n bl ast ul aStage 12.5 13Stage XI V600 700 cel l s i n epi bl astStage 56.5 days post coi t um650 cel l s i n epi bl ast7.5 days post coi t umYol kEarl y embryo Ect oderm M esoderm En

40、doderm Organi zer Yol k Ext raembryoni c t i ssues 1996 Current Bi ol ogysesepxmlmhtexmamnendPSPSendendndneneneyol ksesesepxmpxmhthtlmblblndBLlmlmsmhmhtndSHexmendf bf bmbsehbscscNPSexmlmpxmscscsesehbhbmbmbf bf bmbhbpxmndNlmPSexmendCurrent Biology 1996,Vol 6 No 2Fate maps for the ectodermal layer M a

41、gazi ne105Figure 1Fate maps f or embryos of f our vertebratespeci es,draw n w i t h t hei r normal t opography(see text f or f urt her detai l s).(a)Appearance ofeach embryo bef ore gast rul at i on.Scal e bars:f i sh,500 600 mm;f rog 1 2m m;chi ck,2-3m m;mouse,700 mm.(b)Fate maps at earl ygast rul

42、at i on,drawn i n normal t opography.Embryos are al i gned so t hat t he organi zer i son t he ri ght hand si de of each f ate map.Theorgani zer i s f ound i n t he embryoni c shi el d(SH)of t he zebraf i sh and t he dorsalbl ast opore l i p (BL)of t he f rog embryo;t heprecursor of t he organi zer(

43、t he node)i s f i rstf ound as an i ntegral part of t he earl y primi t i vest reak(PS),t he f i rst morphol ogi cal l andmarkof t he chi ck or mouse embryo.I n represent i ngt he chi ck embryo,separate images of a bl ast o-derm are show n t o i ndi cate germ l ayerprecursors w i t h overl appi ng d

44、i st ri but i ons.(c)Fate maps f or t he ect odermal l ayers at t he endof gast rul at i on(mesodermal and endodermall ayers are not show n due t o spaceconst rai nt s).The anteri or posteri or axes areal i gned hori zontal l y i n t he f i gure,such t hat t het he si te of cel l movement t o t he i

45、 nteri or i s ont he posteri or(ri ght hand si de)of t he map.Byt hi s stage,t he organi zer i s morphol ogi cal l ydi st i ngui shabl e at t he anteri or ext remi t y of t heprimi t i ve st reak(N),as t he H ensen s node(chi ck)or t he node(mouse).Ect odermalt i ssues:ne,neuroect oderm,f b,f orebra

46、i n;mb,mi dbrai n;hb,hi ndbrai n;sc,spi nal cord;se,surf ace ect oderm.M esodermalt i ssues:pxm,paraxi almesoderm;hm,head mesoderm;sm,somi tes;lm,l ateral mesoderm;nd,not ochord;ht,heart;bl,bl ood.Endodermal t i ssues:end,gut endoderm.Ext raembryoni c t i ssues:am,amni on ect oderm;exm,ext raembryon

47、i cmesoderm.Fi shFrogChi ckM ouseAnimal pol eAnimal pol eProximalVegetal pol eVegetal pol eDi st alBl ast ocoelBl ast odermBl ast odermEpi bl astYol k cel l syol k(a)(c)(b)50%epi bol y8000 cel l s i n bl ast oderm90%epi bol ySt age 10+5000 cel l s i n bl ast ul aSt age 12.5 13St age XI V600 700 cel

48、l s i n epi bl astSt age 56.5 days post coi t um650 cel l s i n epi bl ast7.5 days post coi t umYol kEarl y embr yo Ect oderm M esoderm Endoderm Organi zer Yol k Ext raembr yoni c t i ssues 1996 Current Bi ol ogysesepxmlmhtexmamnendPSPSendendndneneneyol ksesesepxmpxmhthtlmblblndBLlmlmsmhmhtndSHexmen

49、df bf bmbsehbscscNPSexmlmpxmscscsesehbhbmbmbf bf bmbhbpxmndNlmPSexmendWheat 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 contribu

50、tions 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-Ep

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