Fertilization and development of mouse oocytes injected with isolated sperm heads.
To determine whether spermatozoa must be structurally intact before microsurgical injection into oocytes for normal fertilization, intact spermatozoa, as well as sperm heads separated from tails by sonication, were individually injected into oocytes. When whole spermatozoa were injected immediately after their immobilization, the majority of the oocytes were fertilized and developed normally. Sonication in the presence or absence of Triton X-100 decapitated more than 95% of spermatozoa. Although all decapitated spermatozoa were diagnosed as "dead" by live/dead sperm staining, separated sperm heads (nuclei) could participate in normal embryo development when injected into the oocytes. The ability of isolated sperm heads (nuclei) to participate in normal embryo development was maintained under cryopreservation conditions that were not suitable for the survival of plasma membrane-intact spermatozoa. These results indicate that 1) spermatozoa do not need to be structurally intact for intracytoplasmic injection, 2) the plasma and acrosomal membranes and all tail components are not essential for normal embryo development, at least in the mouse, and 3) the cryopreservation conditions required for maintenance of the genetic integrity of sperm nuclei are less stringent than those necessary for keeping plasma membrane-intact spermatozoa alive.
Kuretake
S
S
Department of Anatomy and Reproductive Biology, University of Hawaii Medical School, Honolulu 96822, USA.
Kimura
Y
Y
Hoshi
K
K
Yanagimachi
R
R
D000818:Animals; D001755:Blastocyst; D015925:Cryopreservation; D005260:Female; D005306:Fertilization / Q000502:physiology*; D008297:Male; D051379:Mice; D008845:Microinjections; D009028:Morula; D017830:Octoxynol / Q000737:chemistry; D009865:Oocytes / Q000254:growth & development / Q000502:physiology*; D013010:Sonication; D013077:Sperm Head / Q000502:physiology*; D013084:Sperm-Ovum Interactions / Q000502:physiology*; D013089:Spermatocidal Agents / Q000737:chemistry; D013094:Spermatozoa / Q000502:physiology
D013089:Spermatocidal Agents; D017830:Octoxynol