http://genex.hgu.mrc.ac.uk/Atlas/intro.html

Tuesday, March 30, 2010

Wednesday, April 4, 12

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Human 128cell blastocyst before/during hatching

Mouse embryonic stem cells (ES cells)from the inner cell mass, growing in a culture dish

Injection of ES cells into theblastocyst cavity

“zona”

Holding pipetInjection pipet

Oct4 gene expression in epiblast

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Lu, Brennan and Robertson (fig 1)

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from Lu, Brennan and Robertson, 2001

Tuesday, March 30, 2010Wednesday, April 4, 12

Wednesday, April 4, 12

Lu, Brennan and Robertson (fig 5)Gene expression demonstrates the emergence of pattern/breaking of symmetry

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8

Mouse development (Gilbert 6)

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Mouse turning (details are not on the exam)

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10

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11

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12

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Role of the AVE (anterior visceral endoderm) in establishing the anterior-posterior axis of the

mouse embryo

Cylindrical Symmetry Bilateral Symmetry

A P

Cerberus (anti-Nodal)Dkk (anti-Wnt)Lefty (anti-Nodal)

Otx2Anterior neural ectoderm

Nodal, Vg1,Cripto (co-receptorFor Nodal and Vg1)Endo-mesoderminduction

Day 5.25 Day 5.5 Day 6.25

orhypoblast

Endo-mesoderm induction,Primitive streakNodal, wnt, brachyury, fgf expressed

Epiblastectoderm

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Mouse transgenesis (insertion of a foreign gene into the mouse genome)

Gene for red fluorescent protein (RFP)

Gene for green fluorescentprotein (GFP) from jellyfish

(e.g., GFPwith promoter)

FOSTERMOTHER

(2x)

(random)

non-homologous integration

Green ES cells

Dr. MarioCapecchi-1984Univ. of Utah

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Gene knockoutby homologous recombination

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Knockout of the lefty gene illustrates the need for a negative feedback on nodal expression

Mouse Lefty2 and Zebrafish Antivin Are Feedback Inhibitors of Nodal Signaling during Vertebrate Gastrulation

1 Chikara Meno1, 3, Kira Gritsman4, 3, Sachiko Ohishi2, Yasuhisa Ohfuji2, Elizabeth Heckscher4, Kyoko Mochida2, Akihiko Shimono3, 2, Hisato Kondoh3, William S Talbot4, Elizabeth J Robertson5, Alexander F Schier4, 1, Hiroshi Hamada1, 2, 1

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