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Antibody staining (Lecture 3, p.4)

Antibody staining (Lecture 3, p.4). In situ hybridization (Lecture 4 p.4). mRNA depletion (“knockdown”) methods (Lecture 3, p.4). Wnt11 protein. Outside cell. Inside cell. b -catenin mRNA. Axin. b -catenin builds up. b -catenin. Wnt is present.

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Antibody staining (Lecture 3, p.4)

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  1. Antibody staining (Lecture 3, p.4)

  2. In situ hybridization (Lecture 4 p.4)

  3. mRNA depletion (“knockdown”) methods (Lecture 3, p.4)

  4. Wnt11 protein Outside cell Inside cell b-catenin mRNA Axin b-catenin builds up b-catenin Wnt is present. b-catenin protein is not degraded. It builds up, binds to Tcf/LEF and enters the nucleus. Wnt is absent. b-catenin mRNA is continuously translated and the protein is continuously degraded.

  5. Wnt, beta-catenin, and organizer formation * * * * * * *

  6. Endo-mesoderm induction. Pieter Nieuwkoop 1968

  7. (Nieuwkoop center) (Nieuwkoop center)

  8. Nodal signaling pathway(belongs to TGF-beta family of pathways) phospho-Smad2

  9. A dominant negative form of the Nodal (or Bmp) receptor can block Nodal (or Bmp) signaling

  10. Deplete and replace vegT mRNA vegT depleted No mesoderm or endoderm, No blastopore vegT depleted and restored vegT mRNA restored by injection into vegetal hemisphere

  11. VegT depleted embryos: rescue with mRNA for xnr1, 5, 6, or derriere, injected into vegetal pole cells xnr1 Normal late blastula: xnr1 expressed in vegetal cells derriere rescue xnr1 rescue xnr5, xnr6 rescue

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