Fertilization. Animal hemisphere. Sperm entry point
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1 Fertilization Animal hemisphere Sperm entry point Establishes the dorsal/ventral axis Ventral side sperm entry Dorsal side gray crescent Organized by sperm centriole
2 Cleavage Unequal radial holoblastic cleavage Yolk retards division of vegetal pole Second cleavage starts before first is finished
3 Early Cleavage
4 Gray Crescent Opposite to sperm entry Results from rotation of cortical layer of oocyte
5 Cleavage
6 EP-cadherin Cell adhesion molecule Helps hold together blastomeres Translated from mrna deposited in oocyte
7 Anti-sense RNA Used as tool to remove mrna from cell Complement of target mrna Base pairs with target mrna Inactive Destroyed
8 EP-cadherin necessary for cell adhesion Oocyte was treated with anti-sense RNA's against EP-cadherin mrna
9 Mid-blastula Transition Genome activated Embryonic DNA transcribed Cell blastomeres become mobile What triggers MBT? Factors deposited in egg reduced by presence of new chromatin Once threshold level is reached MBT occurs
10 Demethylation Methylation of gene promoters inactives genes Most sperm and oocyte genes Demethylation removes methane groups and allows genes to be transcribed
11 Early Fate Map
12 Cell movements during gastrulation
13 Dorsal blastopore lip
14 Cell from dorsal marginal zone for blastopore groove
15 Involution at blastopore
16 Blastopore formation
17 Expression of paraxial protocadherin
18 Epiboly of Ectoderm
19 Amphibian Axis Animal hemispheres will become ectoderm Vegetal hemispheres will become endoderm Equatorial region will become mesoderm
20 Regulative Development A blastomere has greater potential than its normal cell fate Blastomere fate determined by neighboring cells
21 Spemenn's Experiment Early nuclei are equivalent.
22 Spemann Blastomeres equivalent when they both have gray crescent. Gray crescent necessary for dorsal structures
23 Spemann transplantation experiments Early gastrula cell fate not determined Late gastrula cell fate determined
24
25
26 The Organizer Dorsal lip induces formation of dorsal structures.
27 Vegetal Cell Transplantation
28 Vegetal Cell Transplatation
29 The Organizer Diffusible factor found in dorsal lip cells It itself is induced by underlying vegetal cells Nieuwkoop center beta catenin Functions as a transcription factor and cadherin binding protein
30 Beta-catenin activity in Sea Urchins Determines mesoderm Found in micromeres and veg 2 layer Found in nucleus when active
31 Beta-catenin controlled by Wnt Wnt turns on beta-catenin activity Allows beta-catenin to enter nucleus and act like a transcription factor
32 Wnt Pathway
33 10.2 Reorganization of cytoplasm in the newly fertilized frog egg (Part 1)
34 Beta-catenin localization How does beta-catenin become localized to dorsal side? Beta-catenin evenly distributed before fertilization. Dsh and GBP translocated to dorsal side by kinesin Rides of microtubule tracks laid down after fertilization during cortical rotation Dorsal surface enriched in Dsh and GBP Dsh and GBP inhibit GSK3 Beta-catenin degrades on ventral side and is stabilized on dorsal side
35 Beta-catenin activates other genes Turns on goosecoid gene in organizer
36 Goosecoid mrna can induce new axis Activates involution Determines dorsal mesoderm Represses Wnt8 Important in brain formation
37 VegT and Veg1 Present in vegetal cortical region Genes required for endoderm and mesoderm formation VegT antisence RNA causes epidermis only
38 Xnr Gradient High Xnr Organizer, goosecoid Med Xnr Lateral mesoderm Low Xnr > high BMP4 + Xwnt8 Ventral mesoderm
39 Neural ectoderm induced by soluble factor? Controlled by goosecoid and beta-catenin?
40 Induction of ectopic neural structures
41 Neural structures default epidermis induced by BMP's BMP bone morphogenic protein Induce ectoderm to become epidermis Organizer secretes factors that block BMP from acting
42 BMP inhibitors Noggin, Chordin and Follistatin Diffusable proteins Induced dorsal ectoderm to become neural Dorsalizes mesoderm cells Inhibits BMP's Found in dorsal lip then in the notochord
43 Noggin mrna rescues Dorsal Structures
44 Localization of Noggin
45 Localization of Chordin mrna
46 BMP Gradient Controls Dorsal/Ventral Axis
47 How is the ant/pos axis determined? During neurulation beta-catenin forms a gradient Greatest concentration at organizer Becomes anterior end
48 Wnt Gradient Controls Anterior/Posterior Axis
49
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