Table 2. Pronuclear Microinjection Experiments.
Donor zygotes were generated by ICSI and acceptor parthenotes were generated by activation of oocytes, as described in Methods. Male pronuclei were extracted from zygotes at indicated time points after fertilization and injected into acceptor oocytes at indicated times after activation. Age of the oocytes reflects time from the moment of sperm injection (Exp: ICSI) or moment when the oocytes were placed in SrCl2 containing medium to activate (Exp: P1, P2 and 3–7), and at which the oocytes were placed into media containing BrdU. Age of the transferred pronuclei indicates the time from the moment of sperm injection of ICSI to create the donor zygotes (Exp: 3–7). Oocytes were incubated in BrdU for 30 min (Exp. ICSI, P1, P2, 3, 4, and 6) for 3 hr (Exp. 5) or for 4 hr (Exp. 7), then fixed.
| Exp. ID | Age of Trans’d. PN (hr) |
Age of Oocyte (hr) |
No. Oocytes |
BrdU Positive | |||
|---|---|---|---|---|---|---|---|
| no PN | 1 PN (Trans’d., ♂) |
1 PN (Acceptor, ♀) |
2 PN (♂ + ♀) | ||||
| Controls | |||||||
| ICSI | NA | 7.0 | 22 | 3 (13.6%) | 0 | 1 (4.5%) | 18 (81.8%) |
| P1 | NA | 4.5 | 20 | 20 (100%) | NA | 0 | NA |
| P2 | NA | 7.0 | 28 | 1 (3.6%) | NA | 27 (96.4%) | NA |
| Tested groups | |||||||
| 3 | 7 | 7 | 47 | 0 | 0 | 10 (21%) | 37 (79%) |
| 4 | 4.5 | 7 | 23 | 0 | 0 | 23 (100%) | 0 |
| 5 | 4.5 | 7 | 27 | 0 | 0 | 2 (7.4%) | 25 (92.6%) |
| 6 | 7 | 3 | 37 | 13 (35%) | 24 (65%) | 0 | 0 |
| 7 | 7 | 3 | 21 | 1 (5%) | 1 (5%) | 0 | 19 (90%) |