Bovine embryo development following ICSI: effect of activation, sperm capacitation and pre-treatment with dithiothreitol.
The development of bovine embryos obtained by intracytoplasmic sperm injection (ICSI) was studied in relation to various treatments applied to the sperm and to the early embryo. We investigated the effect of different activation protocols on ICSI-embryos and the influence of sperm capacitation with heparin and D-penicillamine, hypotaurine, and epinephrine (PHE) prior to ICSI. Finally, we studied the effect of dithiothreitol (DTT) pre-treatment of sperm or of injected oocytes. The activation of ICSI-embryos by ionomycin (Io)-cycloheximide (CHX) and sperm pre-treatment with heparin in combination with PHE did not increase the developmental capacity of ICSI-embryos. By contrast, the treatment of injected oocytes with 2 mM DTT resulted in increased cleavage and blastocyst rates in the group of non-activated embryos and in acceleration of blastocyst development in the group of activated embryos. Similarly, pre-treatment of sperm with DTT, followed by ICSI and activation, determined an increase of embryo development on Day 7 although the total number of blastocysts recorded on Day 8 was not different from untreated controls. The transfer of 11 ICSI-blastocysts, produced without activation, in six recipients gave rise to two pregnancies of which one went to term with the birth of an healthy calf.
D000818:Animals; D002417:Cattle / Q000196:embryology*; D003513:Cycloheximide / Q000494:pharmacology; D004229:Dithiothreitol / Q000008:administration & dosage*; D004622:Embryo, Mammalian / Q000187:drug effects; D005314:Embryonic and Fetal Development / Q000187:drug effects*; D004837:Epinephrine / Q000494:pharmacology; D005260:Female; D006493:Heparin / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D008297:Male; D009865:Oocytes / Q000187:drug effects; D010396:Penicillamine / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D011256:Pregnancy Outcome / Q000662:veterinary; D012732:Sex Determination Analysis / Q000662:veterinary; D013075:Sperm Capacitation / Q000187:drug effects; D020554:Sperm Injections, Intracytoplasmic / Q000662:veterinary*; D013094:Spermatozoa / Q000187:drug effects; D013654:Taurine / Q000031:analogs & derivatives* / Q000494:pharmacology
D013654:Taurine; D015759:Ionomycin; C003949:hypotaurine; D006493:Heparin; D003513:Cycloheximide; D010396:Penicillamine; D004229:Dithiothreitol; D004837:Epinephrine