intromcb.berkeley.edu/courses/mcb141/2012/lecture/richard...mouse development (gilbert 6) wednesday,...

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

Tuesday, March 30, 2010

Wednesday, April 4, 12

Wednesday, April 4, 12

Wednesday, April 4, 12

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

Wednesday, April 4, 12

Lu, Brennan and Robertson (fig 1)

Wednesday, April 4, 12

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

Wednesday, April 4, 12

8

Mouse development (Gilbert 6)

Wednesday, April 4, 12

Mouse turning (details are not on the exam)

Wednesday, April 4, 12

10

Wednesday, April 4, 12

11

Wednesday, April 4, 12

12

Wednesday, April 4, 12

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

Wednesday, April 4, 12

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

Wednesday, April 4, 12

Gene knockoutby homologous recombination

Wednesday, April 4, 12

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