1. Cleavage: cutting up the embryo cleavage patterns: why cleavage is important for development. cytokinesis mechanisms: the role of the cytoskeleton, and variation in how membrane is added. regulation mechanisms: the role of the cyclins and mpf: experiments cleavage types: the problem with yolk

2. Mitosis lite. MPF = cyclin B + cyclin-dependent kinase. Cdc2 phosphorylates proteins involved directly in karyo- and cytokinesis. MPF is necessary and sufficient for progression through the cell cycle

3. Experimental test of MPF function in frogs. early embryos incubated in cycloheximide stop cleaving. inject MPF and cleavage resumes. purified MPF Embryo 1. cycloheximide inhibits protein synthesis

4. How are cleavages accomplished? Different molecular mechanisms of cleavage (contractile ring vs. addition of vesicles) actin: bluetubulin: orange photo: G. von Dassow, Center for Cell Dynamics

5. Cleavage type: the problem of yolk Different animals cleave in different ways:

6. How many types of cleavage? 1. (i) Holoblastic (complete) with Isolecithal eggs Observed in eggs with low amounts of yolk. Yolk homogeneously distributed in egg and blastomeres are often the same size. Cleavage can be divided into two categories:

7. How many ways to cleave? 1. (ii) Holoblastic with mesolecithal eggs. Yolk is distributed heterogeneously (concentrated in the vegetal pole). Mesolecithal (intermediate amount of yolk). Cleavage furrow can still penetrate the yolk, but vegetal cells are bigger than animal cells b/c at any given moment in time have undergone fewer cell divisions.

8. How many ways to cleave? 2. (i) Meroblastic: incomplete cleavage with telolecithal yolk distribution. Squid Large yolk contribution precludes complete cleavage Fish Also called discoidal cleavage as there is a disc of cytoplasm on a massive yolk sac.

9. 2. Meroblastic (incomplete) cleavage 2.(ii) Meroblastic: incomplete cleavage with centrolecithal yolk distribution. Many arthropods have this type of cleavage. narrow band of cytoplasm on the outer surface of the egg yolky center cytoplasm

10. In fruit flies karyo- and cytokinesis are uncoupled for the first 12 mitoses. Fruit fly zygote egg plasma membrane Fly karyokinesis (nuclear cleavage) occurs in a syncitium This pattern is thought to have evolved to maximize speed of embryonic development. only seen among holometabolous insects

11. Still other ways of dealing with yolk. gastropods and other animals (e.g. annelids) form polar lobes, generating a quasi-3 cell stage called the trefoil stage. Gastropod Scallop Trefoil

12. Cleavage patterns cleavage planes are: (i) the principal mechanisms by which maternal determinants are sequestered into particular lineages (cytoplasmic determinants) and cleavage is not just about dividing up the embryo: its about dividing it up in a very specific way. (ii) the means by which intercellular interactions are determined

13. In most animals cleavage patterns are not random. The plane of cleavage is always perpendicular to the spindle. Therefore, the orientation of the spindle determines the pattern of cleavage. What determines the orientation of the spindle?

14. The anatomy of an egg Vegetal hemisphere Animal hemisphere Equator Cortex Vegetal pole Animal pole Germinal vesicle (nucleus) Plasma membrane Cytoplasm Maternal axis The animal-vegetal axis (also called the maternal axis) is the reference point for distinguishing among cleavage patterns.

15. Cleavage patterns Radial (biradial): the mitotic spindle is oriented at right angles to the maternal axis Can broadly be divided into two categories, based upon the orientation of the mitotic spindles to the maternal axis. Spiral (duet): the mitotic spindle is oriented at oblique angles to the maternal axis

16. The difference between radial cleavage and spiral cleavage is the orientation of the mitotic spindle to the maternal (A/V) axis. AVAV

17. Radial and Spiral Cleavage Radial: Deuterostomes Spiral: Protostomes

18. Biradial: radial + left/right symmetry established by first cleavage. L/R axis Some modified cleavages: Biradial and Duet

19. Duet Cleavage Acoel flatworms (Phylum Platyhelmintha) The difference between spiral and duet is that the micromeres arise one cleavage earlier (at the four instead of eight cell stage)

20. Why does cleavage plane matter? The plane of cleavage has importance consequences for development. movement of the ABa and ABp relative to P2 reverses the polarity of the worm intercellular communication requires precise orientation of cleavage planes.