intromcb.berkeley.edu/courses/mcb141/2012/lecture/richard...mouse development (gilbert 6) wednesday,...
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http://genex.hgu.mrc.ac.uk/Atlas/intro.html
Tuesday, March 30, 2010
Wednesday, April 4, 12
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Wednesday, April 4, 12
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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)
Wednesday, April 4, 12
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from Lu, Brennan and Robertson, 2001
Tuesday, March 30, 2010Wednesday, April 4, 12
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Wednesday, April 4, 12
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Lu, Brennan and Robertson (fig 5)Gene expression demonstrates the emergence of pattern/breaking of symmetry
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Mouse development (Gilbert 6)
Wednesday, April 4, 12
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Mouse turning (details are not on the exam)
Wednesday, April 4, 12
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Wednesday, April 4, 12
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Wednesday, April 4, 12
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Wednesday, April 4, 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
Wednesday, April 4, 12