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Textbook;  Principles of Development, Lewis Wolpert and Cheryl Tickle. Review papers; PowerPoint Presentation
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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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slide1

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

slide3

Overview of Lectures

Cell Fate

Positional information

Anterior (Head)

Right

Dorsal (Back)

Ventral (Front)

Left

Posterior (Tail)

slide4

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
slide5

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.
slide6

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

slide10

Germ layers, Ectoderm, Mesoderm, and Endoderm, give rise to all tissues

of the developing embryo

Blastocyst

slide11

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.
slide12

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.
slide13

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
slide14

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
slide15

Epigenesis vs Preformation

Nicolas Hartsoeker, 1695

slide16

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’.
slide17

Regulated development in mouse embryos

Donor

Recipient

2-cell

embryo

Tarkowski, (1959)

Nature 184, p1286-7

slide18

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

slide19

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