|
2-cell stage
|
Arrest of DNA synthesis for 2 h with aphidicolin, 2 μg/ml
|
No combs formed (0/30)
|
DNA synthesis is essential for comb formation.
|
|
16-cell stage
|
|
32-cell stage
|
|
60-cell stage
|
27/30 (90%) formed combs, comb formation was delayed by 2 h
|
|
4 hpf
|
29/30 (97%) formed combs, comb formation was delayed by 2 h
|
|
5 hpf
|
29/30 (97%) formed combs, comb formation was delayed by 2 h
|
|
60-cell stage and 4 hpf
|
Arrest of DNA synthesis and cytokinesis with aphidicolin, 2 µg/ml, plus cytochalasin B, 1 to 2 μg/ml
|
Immediately arrested, single nuclei observed, neither combs nor photoprotein were formed in any embryo (0/30)
|
DNA synthesis is essential for comb formation even in cleavage-arrested embryos.
|
|
1- to 2-cell stage
|
Blocking of transcription with actinomycin D, 10 μg/ml
|
0/30 gastrulated, development arrests at 32- to 60-cell stage
|
Zygotic transcription begins around the 32- to 60-cell stage and the specification of the combs requires more than only maternal RNAs.
|
|
4- to 8-cell stage
|
0/30 gastrulated, development arrests at 32- to 60-cell stage
|
|
16- to 32-cell stage
|
10/30 (33%) gastrulate, 20/30 (67%) arrest at 32- to 60-cell stage
|
|
60-cell stage
|
28/30 (93%) gastrulate but no pharynx is formed, 2/30 (7%) do not gastrulate
|
|
5 hpf (post-gastrulation)
|
No pharynx is formed
|
|
2-cell stage
|
Arrest of translation with puromycin, 125 μg/ml, 25 μg/ml
|
0/30 formed combs
|
Comb protein is translated prior to 6 hpf, comb formation requires protein that is translated in the embryo and not maternally loaded.
|
|
16-cell stage
|
0/30 formed combs
|
|
60-cell stage
|
0/30 formed combs
|
|
6 hpf (post-gastrulation)
|
32/30 (97%) formed combs
|
