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. 2012 Aug 6;198(3):357–370. doi: 10.1083/jcb.201202135

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© 2012 Courtois et al.

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Figure 3. — Progressive transition from acentrosomal to centrosomal spindle formation during mouse preimplantation development. (A and B) Live imaging of mouse embryos during the second division (A) and the fourth division (B) at prophase, prometaphase, and metaphase. Note the small microtubule asters (presumably MTOCs; yellow arrowheads) in two-cell and eight-cell stage embryos before NEBD. Red arrowheads mark multipoles of the spindle at prometaphase. Z-projected images of 3-µm confocal sections showing microtubules (EGFP-MAP4; gray) and DNA (H2B-mRFP1). Time is shown in hours and minutes after hCG. (C–E) Immunostaining of embryos fixed during the second division (showing one of the two cells; C), the fourth division (D), and at 113 h after hCG (E4.5; E) at prophase, prometaphase, and metaphase and stained for DNA, microtubules, and pericentrin. In prometaphase at the two-cell and eight-cell stages, several small MTOCs are visible (arrowheads), whereas only two bright MTOCs (arrows) before and after NEBD are seen at E4.5. All pictures are projected images of 0.38-µm stacks. Insets in E represent a zoom of the boxes. Bars: (A–E, main images) 10 µm; (E, insets) 5 µm.