Intracytoplasmic sperm injection in the mouse |
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Authors: | Ron-El R; Liu J; Nagy Z; Joris H; Van den Abbeel E; Van Steirteghem A |
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Institution: | 1Department of Obstetrics and Gynecology, Assaf Harofeh Medical Center Sackler School of Medicine, Tel Aviv University Tel Aviv, Israel
2Center for Reproductive Medicine, Brussels Free University Laarbeeklaan 101, 1090 Brussels, Belgium |
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Abstract: | Intracytoplasmic sperm injection (ICSI) into mouse oocytes involvesa very low survival rate. This study was designed to determinewhy ICSI frequently fails in mice. Metaphase II oocytes wereobtained from superovulated 46 week old F1 hybrid mice.Spermatozoa were retrieved from the epididymis of 1214week old F1 hybrid mice. The spiked microinjection pipette usedto inject a spermatozoon into the ooplasm had outer and innerdiameters of 10 and 8 µm respectively. The oocytes usedin the first part of the study were not activated (group 1).Some oocytes were incubated with calcium ionophore for 5 min(group 2). The injected oocytes were evaluated 6, 20, 48 and72 h after injection. A total of 143 eggs in each group underwentICSI. In group 1, sperm heads escaped into the perivitellinespace. In all, 63 (47%) of the remaining oocytes were damagedduring the injection or had degenerated by the first evaluation.The survival rate was 53%, but fertilization did not occur.In group 2, 31 oocytes (22%) were damaged during microinjectionor soon degenerated. Two oocytes underwent accidental subzonalinsemination. Six oocytes were fertilized (4.2%) among the 78%of survivors. After injection, the sperm tail was found in thecytoplasm (27 and 31% in groups 1 and 2 respectively), the perivitellinespace (45% in both groups) or protruding through the zona pellucida(28 and 23% respectively). More oocytes degenerated when thetail remained in the cytoplasm, i.e. 78% in group 1 and 36%in group 2. |
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Keywords: | assisted fertilization/intracytoplasmic injection/microinsemination |
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