Principles of embryonic patterning
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1 Principles of embryonic patterning Hilde Van de Velde Centre for Reproductive Medicine Centre for Medical Genetics Department of Embryology and Genetics ESHRE2010, October 9
2 Outline Reproductive Biology Lessons from animal models Totipotency (Pre-)patterning and destiny Lessons from the human embryo Totipotency and differentiation Conclusions
3 Outline Reproductive Biology Lessons from animal models Totipotency (Pre-)patterning and destiny Lessons from the human embryo Totipotency and differentiation Conclusions
4 Totipotency Life starts with... the totipotent cell The totipotent cell is... able to develop into fertile offspring The ultimate totipotent cell is... the zygote Embryonic patterning?
5 Developmental biology: Echinodermata Sea urchin Direction is reversible Blastomeres are totipotent, plastic Regulative development
6 Developmental biology: Invertebrates,Tunicata Sea squirt Direction is irreversible: commitment, destiny or fate Restricted development
7 Developmental Biology: human monozygotic twinning
8 Lessons from the mouse embryo Pre-patterning in the zygote Gardner et al. 1997, 2001 Em Em V A Abem Abem 2nd Pb is fixed
9 Lessons from the mouse embryo Piotrowska et al the 1st dividing blastomere contributes more to the Em part pre-patterning at the 2-cell stage manipulations, in vitro experiments ME
10 Lessons from the mouse embryo Piotrowska et al. et al. 2001
11 Lessons from the mouse embryo Torres-Padilla et al V blastomere is less stained for H3R26me CARM1 (H3-specific arginine transferase) 88.5% ICM cells derived from injected blastomere (overexpression Nanog)
12 Lessons from the mouse embryo Regulative development The dance of the embryo (time lapse) Kurotaki et al ZP (extrinsic factor) induced cavity 2nd Pb is not fixed, embryo rotates in the ZP In vivo experiments: photoconversion No pre-patterning
13 Lessons from the mouse embryo Inside-outside hypothesis Tarkowsky and Wroblewska 1967 IN ICM OUT TE
14 Lessons from the mouse embryo Potency of IN and OUT blastomeres Suwinska et al cell stage Compacted puppies puppies 32-cell stage Early blastocyst ICM+TE, no puppies TE direction is reversible at compaction but irreversible at blastulation
15 Lessons from mouse embryo Regulative development Suwinska et al cell stage Early blastocyst sorting puppies
16 Lessons from the mouse embryo Polarization Cockburn and Rossant, 2010 Compaction: E-cadherin Basolaterally: nuclei; Par1, Lgl Apically: endosomes; actin, apkc, Par3
17 Lessons from the mouse embryo Polarization Johnson and McConnel, 2004; Cockburn and Rossant, 2010
18 Lessons from the mouse embryo Lineage segregation Vallier and Rossant, st lineage TE: Cdx2 ICM: Pou5F1/Nanog 2nd lineage Epi: Nanog PE: Gata4/6
19 Lessons from mouse embryo Stochastic model = regulative development Dietrich and Hiiragi, 2007 Oct-4: uniform and sorted Cdx2 and Nanog: 2 phases (1)Variable (2)Sorted
20 Lessons from the mouse embryo 1st lineage segregation Ralston and Rossant, 2005 Cdx2 downstream of polarization Jedrusik et al. 2008; 2010 Cdx2 mrna is polarized in outside cells
21 Lessons from the mouse embryo 1st lineage segregation Jedrusik et al. 2008; 2010 Cdx2 mrna: upregulation - Greater contribution to TE - More symmetric divisions - apkc locatization dsrna Cdx2: downregulation - Greater contribution to ICM
22 Lessons from the mouse embryo 1st lineage segregation Nishioka et al. 2009; Cockburn and Rossant, 2010 Different cell contact Lat1/2 kinases Apical domain
23 Lessons from the mouse embryo 2nd lineage segregation Chazaud et al Pepper-and salt-distribution Nanog and Gata6 Sorting Epi: Nanog PE: Gata4/6
24 Lessons from mouse embryo Zernicka-Goetz et al Cell position Cell movement according to gene expression pattern Changes in gene expression Apoptosis
25 Outline Reproductive Biology Lessons from animal models Totipotency Pre-patterning and destiny Lessons from the human embryo Totipotency and differentiation Conclusions
26 Lessons from the human embryo Embryonic genome activation (day 2/3) (Braude et al. 1988; Dobson et al. 2004; Cauffman et al. 2005; Cauffman et al. 2006) 1 st differentiation (day 5) TE: differentiated ICM: pluripotent Extra-embryonic endoderm, mesoderm and ectoderm 3 germ layers PGC embryonic stem cells (hesc)
27 Lessons from the human embryo Totipotency Marker for totipotency? When is totipotency lost? When is the 1st differentiation irreversible? Regulative development Totipotency Differentiation
28 Lessons from the human embryo No stemness marker to identify totipotent cells Cauffman et al. 2005; 2006; 2009 Boyer et al hesc NANOG SOX2 POU5F1_iA
29 Lessons from the human embryo Embryo splitting Van de Velde et al Sister 4-cell stage blastomeres are potentially totipotent
30 Lessons from the human embryo Splitting 8-cell stage embryo No totipotent capacity or not enough cells to form an inner cell population?
31 Outline Reproductive Biology Lessons from animal models Totipotency Pre-patterning and destiny Lessons from the human embryo Totipotency and differentiation Conclusions
32 Conclusions Patterning in the mouse Regulative development Polarity and position 1st and 2nd differentiation Cell movement, changes in gene expression, apoptosis Patterning in the human Regulative development
33 Thanks Greet Cauffman Inge liebaers Caroline De Paepe Maria Krivega Martine De Rycke Laetitia Petrussa Karen Sermon Mieke Geens Ileana Mateizel Johan Sterckx Heidi Van Ranst Griet Meersdom An Verloes Martine Vercammen Ewart Kuyck
34 Preimplantation genetic diagnosis De Vos et al Cohort of day 5 SET, 1-cell versus 2-cell biopsy PGD and PGS All embryos resulted from 8-cell stage embryos on day n= n= (control) n=702 P value hcg per ET LBR per ET 46.7% 37.4% 36.3% 22.4% 48.6% 35.0% Only 1 GTE 25.9% 18.2% More than 1 GTE Non-elective SET Elective SET 46.5% 34.1% 55.0% 27.6% 22.1% 35.4%
35 Lessons from the human embryo Two hesc lines of distinct embryos Geens et al VUB_26Quatro: 46 XX mosaic dup(7)(q33qter), del(18)(q23qter) VUB_27Patru: 46 XY normal At least one blastomere is pluripotent at the 4-cell stage Day 4 Day 5 Passage 1 day 2
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