Videos of Cells and Embryos

 
 

The extremely asymmetric divisions of oocyte meiosis – by which oocytes become eggs – follow the central rule of animal cytokinesis: the cleavage furrow bisects the spindle.  It’s just that in oocyte meiosis, the spindle is small and directly associated with the plasma membrane.  The tiny polar bodies that result are dead-end cells containing throwaway copies of the maternal genome, but accuracy is at least as important as in normal cell division because partitioning errors would affect the egg and the embryo that develops from it.

This starfish oocyte expresses a probe for active Rho.  Immediately after the metaphase/anaphase transition, the entire oocyte surface experiences a short-lived wave of Rho activation, which heightens and “breaks” around the meiotic spindle, culminating in a bright zone of active Rho equivalent to the Rho zone in mitotic cytokinesis.  Rho activity in turn prompts actin assembly and myosin recruitment to mediate cell surface contractility, which is what pinches off the polar body.  In most oocytes Rho activation proceeds from vegetal pole to animal (where, by definition, the meiotic spindle is located); in this particular case, the wave travels too rapidly to be obviously directional unless one steps through the movie frame by frame (see the third movie below for a more obvious vegetal-to-animal wave).

Rho activity during polar body emission in starfish

November 19, 2013

Species:

Patiria miniata

Frame rate:

20 sec/frame @ 15 fps = 250-fold time-lapse

Points of interest:

polar body; meiosis; asymmetric cell division; cytokinesis

What’s glowing:

Expressing GFP-rhotekin rGBD, which binds active Rho

Optics:

Olympus FluoView 1000 laser scanning confocal; 60x; projection of 10 1-µm sections.

Collected by:

George von Dassow

More like this:

Get larger versions of two of these two movies: 071911_PmRho-pb1and2_01-lg.mov and 071911_PmRho-pb1and2_02ab-lg.mov

The second movie is nearly the same (it’s even from the same batch of oocytes) but is closer to a true medial section, so the ghostly cloud of the spindle and the maternal pronucleus are visible.

Finally, this third movie is from a different batch of oocytes with a much more pronounced vegetal-to-animal wave.  We don’t know what causes variation in the wave speed or the intensity of the pile-up as it breaks around the spindle, but it seems likely that it’s related to the detailed dynamics of the anaphase drop in cyclin levels associated with the transition from M-phase to C-phase: for some reason it appears that C-phase starts sooner at the vegetal pole, suggesting that cyclin levels fall below some threshold there first, and slightly later pass the same threshold near the spindle.


— text by George von Dassow