5. DEVELOPMENT OF BLOOD
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Transcript of 5. DEVELOPMENT OF BLOOD
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5. DEVELOPMENT OF BLOOD
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Mesonephric tubules
Blood cells
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primitive hematopoiesisventral mesoderm Fish gastrulation movie
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8h embryoICM
2d embryo circulation
scl, gata1/2, lmo2 - transcriptional factors that specify early hematopoietic tissue (gata2 expressed also in ectoderm)RBI – rostral blood islandsICM – inner cell mass
5-somite stage embryo – hematopoiesis starts officially
gata1 promoter-driven GFP = early erythroid lineage
dorsal aorta
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MYELOPOIESIS STARTS IN RBI @ 10-SOMITE STAGEspi1 – marker of myeloid lineage
rostral
caudal
spi1 detected by Fast Red riboprobe
scl – early hematovascular cell fatefli1 – early vascular fatenkx2.5 – heart fate
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7.5 dpc mouse embryo (late gastrulation stage)
8.5 dpc – yolk sac encloses entire embryoblood islands merge to form vascular channels
LacZ driven by -globin promoter (X-gal staining – primitive erythroblasts)
bl - blood islandsec - ectoplacental coneac - amniotic cavityte - trophectodermal - allantoiseem - extraembryonic mesoderm (blue)ve - visceral endoderme - embryonic ectoderm
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AORTA-GONAD MESONEPHROS (AGM) Development of intra-embryonic hemogenic site
fetal liver hemogenic site
aorta
HSC
A- aortaNO- notochordAV- axial veine - endothelial cell
A
lateral plate mesoderm
ICM
Rohon-Beard neurons
RUNX1 riboprobe
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RUNX1 (CBFA2, AML1) is necessary for ‘definitive’ hematopoiesis to start at AGM
Runx1+/-
E11.5: dark- hematopoietic cells
only a few primitive erythrocytes left – likely a carry-over from the ‘primitive’ hematopoiesis
Runx1-/-
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PRIMITIVE vs. DEFINITIVE EXTRAEMBRYONAL vs EMBRYONAL
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BONE MARROW NICHE IS CRITICAL FOR HEMATOPOIESIS
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COLLAGEN TYPE-X DELETION
wt Col X-/-
normal hematopoiesis aplasia
wt
Col X-/-
wt Col X-/-
thymus
spleen
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THE HUNT FOR HEMANGIOBLAST AND ITS HEM- ANGIO-DIFFERENTIATION
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HEMANGIOBLASTscattered among endothelial and HSC in heart field of the gastrula
ventral marginal cells of the heart field, n- notochord
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Endothelial progenitor
HEMANGIOBLAST or HAEMOGENIC EPITHELIA, or BOTH?
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blast colony-forming cell
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Hemangioblast culture
Angiopoietin 1 rec.Integrin alpha 2B
tight adherent structure
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Tie2hic-KIT+CD41_
can generate hematopoietic precursors
endothelial markers endothelia in matrigel
Tie2hic-KIT+CD41_
are present in vivo by FACS
Tie2hic-KIT+CD41_
are present in vivo by IHC
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Runx1-/-
Blast movies 3, 4Runx-/- movie
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TRANSCRIPTIONAL FACTORS IN BLOOD DEVELOPMENT
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GATA-1 - binds DNA via a zinc finger motif
Dracula fish – loss-of-function mutation in GATA1 – impaired erythroid differentiation
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Spadetail/TBX16 DNA binding domain derived from the prototype gene called transcription factor T
impaired erythroid but not myelopoietic differentiation
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Moonshine TIM-family of transcriptional factors
‘primitive’ – survival of HSC ‘definitive’ – cardiomegaly and impaired red cell differentiation
erythrocyte proerythroblast
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WHEN SOMETHING GOES WRONG WITH BLOOD
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ACUTE LYMPHOID LEUKEMIA
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CHRONIC MYELOID LEUKEMIA
bcr
abl
bcr-abl
mice transplanted with patient bone marrow
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