Birth of live mice derived by in vitro fertilization with spermatozoa retrieved up to twenty-four hours after death.
The objective of this study was to determine the viability and fertility of mouse spermatozoa obtained at various postmortem intervals. Male mice were euthanized, and the bodies were kept at approximately 22 degrees C for up to 24 hr. The epididymides were removed and spermatozoa were allowed to swim out into Minimum Essential Medium supplemented with bovine serum albumin. Motility of spermatozoa retrieved within 12 hr postmortem was approximately 60%, whereas motility of those obtained 6 to 12 hr later decreased significantly (P < 0.05). There appeared to be no differences in the percentages of spermatozoa with intact plasma and acrosomal membranes regardless of the time after death. After in vitro fertilization of oocytes with spermatozoa collected immediately after death or at 6, 12, 18, or 24 hr postmortem, the cleavage rates were 81%, 70%, 64%, 34%, and 19%, respectively. Once oocytes were fertilized, more than 65% developed into morulae/blastocysts. Transfer of a total of 166 embryos produced in vitro with postmortem spermatozoa resulted in the birth of 44 live pups (26.5%). Of these 44, 3 live mice were derived by transfer of 11 embryos (27.3%) produced with 24-hr postmortem spermatozoa. Histological examination of the testes and epididymides obtained at various postmortem intervals revealed that degenerative changes of the testes occurred within 6 hr, whereas those of the epididymides were less obvious until 6 hr later. These changes included pyknosis, release of intracellular contents, and disruption of intercellular bridges of the germ cells. This study has demonstrated that spermatozoa recovered from a dead animal as long as 24 hr after death can be used to fertilize oocytes, and that the resulting zygotes can develop into live young.
Songsasen
N
N
Department of Biomedical Sciences, University of Guelph, Ontario, Canada.
Tong
J
J
Leibo
S P
SP
D000177:Acrosome / Q000502:physiology*; D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology; D002417:Cattle; D002470:Cell Survival; D015925:Cryopreservation; D004624:Embryo Transfer; D004822:Epididymis / Q000473:pathology; D005260:Female; D005298:Fertility; D005307:Fertilization in Vitro; D008098:Litter Size; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D009028:Morula / Q000166:cytology / Q000502:physiology; D009865:Oocytes / Q000166:cytology / Q000502:physiology*; D011180:Postmortem Changes; D011555:Pseudopregnancy; D012099:Reproductive Techniques; D012662:Semen Preservation; D012710:Serum Albumin, Bovine; D013081:Sperm Motility; D013094:Spermatozoa / Q000166:cytology* / Q000502:physiology*; D013737:Testis / Q000473:pathology; D013997:Time Factors; D015053:Zygote / Q000166:cytology / Q000502:physiology
D012710:Serum Albumin, Bovine