Table 1.
Short- and Long-term Effects of Heterologous Centrin 3 in Two-Cell Stage Injected Xenopus Embryos
| Pre-MBTdevelopment | Post-MBTdevelopment | ||||||
|---|---|---|---|---|---|---|---|
| Injected solution | N | U | L | N | E | L | Eggs |
| % | % | % | % | % | % | n | |
| Protein injection | |||||||
| HsCen3p (1.5 mg/ml) | 80 | 20 | 0 | 49 | 48 | 3 | 90 |
| HsCen3p (3 mg/ml) | 25 | 75 | 0 | 25 | 75 | 0 | 20 |
| HsCen3p (6 mg/ml) | 30 | 70 | 0 | 10 | 48 | 42 | 91 |
| HsCen3p (12 mg/ml) | 28 | 72 | 0 | 6 | 25 | 69 | 77 |
| Heat-treated HsCen3p | 31 | 69 | 0 | 10 | 77 | 13 | 97 |
| Protease-treated HsCen3p | 95 | 5 | 0 | 58 | 28 | 14 | 56 |
| PBS | 98 | 2 | 0 | 75 | 23 | 2 | 133 |
| RNA injection | |||||||
| WT HsCen3 | 63 | 32 | 5 | 30 | 46 | 24 | 106 |
| Mutant HsCen3 | 38 | 58 | 4 | 6 | 29 | 65 | 103 |
| GFP | 89 | 11 | 0 | 64 | 19 | 17 | 37 |
| Hepes | 94 | 5 | 1 | 64 | 25 | 11 | 79 |
25 ml of native or treated recombinant HsCen3p, or 25 ml of wild-type HsCEN3 (WT), mutant HsCEN3, or GFP RNA at 0.6 μg/ml or control solutions (PBS or Hepes) were injected in one blastomere of two-cell stage embryos. Short-term effects were observed 7 h (protein injection) or 11 h (RNA injection) after fertilization. The effect on blastomere cleavage was scored as normal (N), under-segmented (U), or lysis (L) of the injected half. Long-term effects were observed 22 h after fertilization. Development of the injected side of the embryo was scored as normal (N), participating in exogastrulation of the whole embryo (E), or lysed (L). Note that in the case of RNA injections, pre-MBT development was analyzed 11 h after fertilization to allow RNA translation.