Control of Cleavage Spatial control of cleavage

-karyokinesis & cytokinesis -karyokinesis: definition- mitotic division of nucleus -nucleus position determines type of cleavage, i.e. holoblastic / meroblastic -mitotic spindle & its microtubules (composed of tubulins) Mitotic spindle axis determines plane of cleavage always perpendicular to this axis regardless of location. e.g. equal or unequal divisions Mitotic spindle orientation determines pattern of cleavage, e.g. dextral of sinistral coiling

Genetic control of embryonic cleavage pattern & shell coiling in snail. A) wild type- right-handed (dextral) coil of shell. B) mutant allele- left-handed (sinistral) coil of shell

cytokinesis: definition-division of the cell

- contractile ring of microfilaments (made of actin)

- interaction of astral fibers of centrosomes & plasma membrane required for cytokinesis

Role of microtubules and microfilaments in cell division. (A) Diagram of first-cleavage telophase. The chromosomes are being drawn to the centrioles by microtubules while the cytoplasm is pinched in by the contraction of microfilaments. (B) Localization of actin microfilaments in the cleavage furrow. Fluorescent labeling of the actin microfilaments shows the contractile ring in the first-cleavage furrow (arrowhead) of a telophase sea urchin egg. (C) Fluorescent labeling of tubulin shows the microtubular asters of a sea urchin egg during telophase of first cleavage. (B from Bonder et al. 1988; C from White et al. 1988; photographs courtesy of the authors.)

Mitotic spindle
Centrosomes organize mitotic spindle that leads to formation of cleavage furrow

In holoblastic cleavage, cleavage furrow forms contractile ring

In meroblastic cleavage, cleavage furrows begins as cytoplasmic infoldings Cleavage arrest can be done with cytochalasin which inhibits cleavage by disrupting microfilament formation

Cytokinesis
Occurs in plane perpendicular to axis of mitotic spindle and equidistant between spindle poles

Actin and myosin form contractile ring

Spindle poles become anchored at specific sites in the egg or blastomere cortex and their orientation determines cleavage plane Spindle orientation controlled by material cytoplasmic factors as well as mechanical constraints

Temporal control of cleavage

early divisions are rapid because there is abundant stored mRNA and proteins required for cell divisions (skip G1)

as these stored materials get depleted, embryo cells start transcription and translation
when embryonic machinery begins, it is known as midblastula transition (MBT) internal clock regulated by a biochemical oscillator that regulates cyclin synthesis and MPF generation

MPF (Mitosis or Maturation Promoting Factor)

- Transition from fertilization to cleavage is caused by MPF - all cell divisions are directed by the presence of a complex of molecules called MPF MPF contains 2 subunits: Cyclin B: large subunit, regulates small subunit of MPF, its presence is regulated within all cells in a cyclic fashion. Its presence dictates whether MPF is active or not. It activates cdk. cyclin-dependent kinase (cdk): a molecule that acts on specific target molecules by phosphorylating (or de-phosphorylating) them, activates mitosis by bringing chromatin condensation, nuclear envelope depolymerization & organization of mitotic spindle.

- different forms of both cyclin B and cdk are present at different stages of the cell cycle, creating a complex layering of regulation that can be achieved at each step in the cell cycle

- The cell cycle is regulated by presence of cyclins:

Cyclins degrades and is re-synthesized at particular times in the cell's cycle.
-During S phase (DNA synthesis), after DNA has been

synthesized, -cyclins is synthesized anew during Gap2 (G2), creating active MPF, which activates target molecules that permit the cell to begin the events of mitosis or meiosis

Cell cycles of somatic cells & early blastomeres. A) simple biphasic cell cycle of early amphibian blastomeres- 2 states, S & M. Cyclin allow progression to M (mitosis) while cyclin degradation allows cells to pass into S (synthesis) phase. B) Cell cycle of a typical somatic cell. Cells differentiating is taken out of the cycle and are in an extended G1 phase called G0. Cyclins & respective kinases are responsible for progression through the cell cycle.

In cleavage divisions, the cell passes directly through Gap2 to enter mitosis What is unique about cleavage divisions is that the Gap phases are almost entirely eliminated because: the egg has stored the organelle components, energy sources, etc., that mature cells normally need to stop and synthesize between both mitosis and DNA synthesis the process of DNA synthesis is greatly accelerated in cleavage mitosis the synthesis and degradation of cyclins is speeded up in cleavage stage divisions under direction of yet again another set of stored molecules

Cyclin's presence in the cell is itself regulated by a number of molecules most of which have yet to be identified
they either cause the degradation or synthesis of cyclin The most well known is the protein product of a protooncogene named c-mos, called "cytostatic factor". By regulating its presence, the cell can regulate the presence of cyclin

*Loss of control = uncontrolled cell division, a key feature of cancer cells.

- mutated genes that direct cells to become cancerous are called oncogenes

Special features of MPF regulation during fertilization and cleavage divisions: entry of Ca++ into egg's cytoplasm at fertilization triggers the completion of egg meiosis in the organisms which stop meiosis in the egg pending fertilization. Ca++ does this by regulating the presence of cytostatic factor(s), which in turn regulates the presence of cyclin. Where do all these molecules (cytostatic factor, cyclin and cdk) come from? - some are stored as proteins in the egg, while others must be synthesized anew from mRNAs stored in the egg's cytoplasm as part of the egg's instructional molecule complex.

the events of completion of egg meiosis and the first zygotic mitosis may be directed by protein stored in the egg, but cleavage mitoses rapidly begin to use new proteins synthesized from their mRNA precursors. the Ca++ entry at fertilization also triggers this 1st burst of protein synthesis in the egg after fertilization. mRNAs for many proteins essential for the early steps in cleavage are translated in this burst. different organisms make use of this initial protein synthesis burst to different degrees.

Note that since the mRNAs are already present, this translation can occur in the complete absence of any activation of DNA transcription from the new zygotic genome.

Thank You!