antibody staining (lecture 3, p.4). in situ hybridization (lecture 4 p.4)
TRANSCRIPT
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Antibody staining (Lecture 3, p.4)
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In situ hybridization (Lecture 4 p.4)
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mRNA depletion (“knockdown”) methods (Lecture 3, p.4)
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-catenin
-catenin builds up
Axin
Wnt is absent. -catenin mRNA is continuouslytranslated and the protein is continuously degraded.
Wnt is present. -catenin protein is not degraded. It builds up, binds to Tcf/LEF and enters the nucleus.
Wnt11 protein
Outside cell
Inside cell
-cateninmRNA
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Wnt, beta-catenin, and organizer formation
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Endo-mesoderm induction. Pieter Nieuwkoop 1968
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(Nieuwkoop center)
(Nieuwkoop center)
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Nodal signaling pathway(belongs to TGF-beta family of pathways)
phospho-Smad2
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A dominant negative form of the Nodal (or Bmp)
receptor can block Nodal (or Bmp) signaling
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Deplete and replace vegT mRNAvegT depletedNo mesoderm or endoderm,No blastopore
vegT mRNA restored by injection into vegetal hemisphere
vegT depleted and restored
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VegT depleted embryos: rescue with mRNA for xnr1, 5, 6, or derriere, injected into vegetal pole cells
Normal late blastula:xnr1 expressed in vegetal cells
derriere rescue
xnr5, xnr6 rescue
xnr1 rescue
xnr1