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Textbook;  Principles of Development, Lewis Wolpert and Cheryl Tickle. Review papers;

Reading list. Textbook;  Principles of Development, Lewis Wolpert and Cheryl Tickle. Review papers; Lecture 1 and 2 Alexandre (2001) International Journal of Developmental Biology 45, p457-467 Rossant (2001) Stem Cells 19, p477-82

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Textbook;  Principles of Development, Lewis Wolpert and Cheryl Tickle. Review papers;

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  1. Reading list Textbook;  Principles of Development, Lewis Wolpert and Cheryl Tickle. Review papers; Lecture 1 and 2 Alexandre (2001) International Journal of Developmental Biology 45, p457-467 Rossant (2001) Stem Cells 19, p477-82 Yamanaka et al, (2006). Developmental Dynamics 235, p2301-2314 Katsuyoshi and Hamada, (2012) Development 139, p3-14 Lecture 3 and 4 Arnold and Robertson (2009) Nature reviews Molecular cellular biology, 10, p91-103 Robb and Tam (2004) Seminars in Cell and Developmental biology 15, p43-54 Hayashi et al (2007) Science 316, p394-396. Hashimoto and Hamada (2010) , CurrOpin Genet Dev 20, p433-7 Hanna et al (2010) Cell 143, p508-525. Yamanaka and Blau (2010) Nature 465, p704-712

  2. Model systems for studying vertebrate development

  3. Overview of Lectures Cell Fate Positional information Anterior (Head) Right Dorsal (Back) Ventral (Front) Left Posterior (Tail)

  4. Lecture 1 • Overview of early mammalian development • Fertilisation and parthenogenesis • Mosaic vs regulated development • You should understand • Non-equivalence of maternal and paternal genomes • Mammalian development is highly regulated

  5. In utero development in mouse occurs over 19-21 days • E (embryo stage) = dpc (days post coitum). Most commonly referred to from 0.5 onwards • as mating takes place at night. • Preimplantation development occurs up to E3.5. All other development occurs • postimplantation.

  6. 0 1 2 3 4 Preimplantation Development days Cleavage stages Primitive (primary) endoderm Blastocoel cavity Blastomere Inner cell mass/ Primitive ectoderm Zona pelucida Trophectoderm Activation of embryonic genome

  7. Early Post-implantation Development

  8. Gastrulation and Beyond

  9. Extraembryonic tissues

  10. Germ layers, Ectoderm, Mesoderm, and Endoderm, give rise to all tissues of the developing embryo Blastocyst

  11. Non-equivalence of maternal and paternal genomes • Penetration of cumulus cells • Acrosomal reaction penetrates zona pellucida made up of glycoproteins • Sperm and egg plasma membranes fuse and sperm nucleus enters egg. • Fertilization triggers dramatic release of calcium in the egg, setting in train completion of • female meiosis etc.

  12. Pronuclear Maturation Second polar body Zonapelucida Syngamy Male pronucleus. Female pronucleus. 12 24 0 hr post fertilization Replication initiation M-phase • Maternal and paternal haploid genome remains separate (pronuclei) until first metaphase.

  13. Parthenogenesis Parthenogenetic activation - Genetic background - In vitro manipulation - Pronase/hyalouronidase - Heat shock - Ethanol - Strontium chloride • Oocytes can be activated in the absence of fertilization, leading to parthenogenetic development • Parthenogenetic embryos have limited viability, contrasting with other model organisms • Limited viability suggests either that sperm/fertilization confers essential properties for development or • that maternal genome alone is incapable of supporting development

  14. Recipient zygote Donor zygote Non-equivalent contribution of maternal and paternal genomes ? Barton, Surani , Norris (1984) Nature 311, p374-6 McGrath and Solter, (1984) Cell 37, p179-183 • Gynogenetic embryos have retarded growth/development of extraembryonic tissues • Androgenetic embryos have retarded growth/development of embryonic tissues

  15. Epigenesis vs Preformation Nicolas Hartsoeker, 1695

  16. Mosaic and Regulated development • Roux (1888) shows ‘mosaic development’ of frog embryo following ablation of one cell in • two-cell embryo – formation of ‘half’ embryo. • Driesch (1895) finds opposite is true for sea urchin, normal albeit smaller embryo develops • from one of two cells – ‘regulated development’.

  17. Regulated development in mouse embryos Donor Recipient 2-cell embryo Tarkowski, (1959) Nature 184, p1286-7

  18. 8-cell embryos Remove zona pellucida Aggregate in dish Culture in vitro Transfer to foster mother Chimeric blastocyst Chimeric progeny Chimeras from aggregaton of 8-cell stage embryos Tarkowski (1961) Nature 190, 857-860

  19. Chimeras from transfer of ICM cells Gardner (1968), Nature 220, p596-7 • Gardner later demonstrated this for ICM cells of the blastocyst stage embryo. • In these experiments ICM cells did not contribute to trophectoderm or primitive endoerm lineages

  20. End lecture 1

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