Premium
Subzonal insemination with a single spermatozoon using manipulation assisted sperm adhesion onto the ooplasmic membrane in mouse ova
Author(s) -
Kobayashi Kazuhiko,
Okuyama Manabu,
Fujimoto Goro,
Ogawa Shyoso,
Rothman Cappy M.,
Hill David L.
Publication year - 1992
Publication title -
molecular reproduction and development
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.745
H-Index - 105
eISSN - 1098-2795
pISSN - 1040-452X
DOI - 10.1002/mrd.1080310310
Subject(s) - sperm , spermatozoon , human fertilization , capacitation , zona pellucida , andrology , biology , insemination , population , adhesion , oocyte , microbiology and biotechnology , anatomy , embryo , chemistry , medicine , environmental health , organic chemistry
Abstract A simple and successful method of microinjection of a single spermatozoon under the zona pellucida of a mouse oocyte has been developed. A characteristic of this method is that the tip of the sperm injection needle pierces the zona pellucida without touching the ooplasmic membrane. All the ova (277) used for this series of experiments had normal morphology after the injection procedure. Spermatozoa preincubated in culture medium for capacitation and those treated with ionophore A23187 for induction of acrosome reaction were used. In combination with some of these injections, a manipulation assisting the adhesion of the sperm head onto the ooplasmic membrane was employed. The fertilization rate (67.3%) of the ova injected with the ionophore‐treated sperm using the sperm‐adhesion treatment was significantly higher ( P <0.005) than that obtained by the injection of the preincubated sperm without applying the adhesion treatment (23.6%). All three of the recipients that received the 24 fertilized ova became pregnant and gave birth to 11 off‐spring (45.8%). The inseminations performed with the sperm‐adhesion treatment using the immotile sperm from the preincubated population and/or those from the ionophore‐treated population did not result in fertilization in any case. These results suggest that the fertilization rate of subzonal insemination with motile ionophore‐treated sperm can be improved by applying the sperm‐adhesion treatment and that sperm motility might be involved in the establishment of fertilization, even after the adhesion of the sperm head with the mouse ovum membrane.