title: A comparison between in vitro fertilization and microinjection of immobilized spermatozoa from bulls producing spermatozoa with defects.
abstract: The objectives of this study were to compare the fertilization rate of bovine in vitro matured oocytes by in vitro fertilization (IVF) and by microinjection of a single spermatozoon (MI) and to relate these rates with fertility reported for these bulls in artificial breeding. Bull A (Holstein) had a nonreturn rate of 75%. Semen from this bull is routinely used in our standard IVF procedure. Bull B (Ayrshire), used regularly in artificial breeding and related to bull D, had a nonreturn rate of 69.2%. Bull C (Brown Swiss), with a chromosomal translocation and trisomy, achieved a nonreturn rate of 42%. Bull D (Ayrshire) produced nonmotile spermatozoa (SPZ) and had an abnormality described as "tail stump defect." No pregnancies sired by bull D have been reported. Oocytes were either fertilized in vitro by capacitated SPZ or by microinjection of a single immobilized SPZ into the ooplasm. SPZ were treated with 0.1 microM A23187 and used for IVF. For microinjection SPZ were cocultured for 5 h with bovine oviduct epithelial cells (BOEC) and then immobilized by freezing and thawing twice without cryoprotectant. A single batch of killed SPZ (stored at -25 degrees C) was used for all microinjections. All oocytes were cultured in Medium 199 for 22 h at 39 degrees C and subsequently fixed, stained, and examined for evidence of fertilization (i.e., female and male pronucleus formation, SPZ decondensation). Fertilization rates following IVF with semen from bulls A, B, C, and D were 80%, 54%, 1%, and 2%, and following microinjection were 39%, 22%, 21%, and 34%, respectively.
authors: Department of Animal Science, Cornell University, Ithaca, New York 14853-4801.
authors: W
authors: W
authors: Heuwieser
authors: X
authors: X
authors: Yang
authors: S
authors: S
authors: Jiang
authors: R H
authors: RH
authors: Foote
mesh_terms: D000818:Animals; D002417:Cattle; D005069:Evaluation Studies as Topic; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods*; D066298:In Vitro Techniques; D007248:Infertility, Male / Q000473:pathology / Q000503:physiopathology; D008297:Male; D008845:Microinjections; D009865:Oocytes / Q000502:physiology; D013081:Sperm Motility; D013094:Spermatozoa / Q000002:abnormalities / Q000502:physiology