title: Effects of sperm pretreatment and embryo activation methods on the development of bovine embryos produced by intracytoplasmic sperm injection.
abstract: The aim of the study was to examine the effects of different embryo activation methods and sperm pretreatments on the activation and development of bovine embryos produced by intracytoplasmic sperm injection (ICSI). Four activation agents, i.e., calcium ionophore (A23187), ionomycin (Ion), electric pulse (EP) and ethanol (Eth) were used in various combinations to activate bovine ICSI embryos. The normal fertilization rate was similar in bovine ICSI embryos activated by A23187+Eth, Ion+Eth, Ion+EP+Eth, and 2-Ion (Ion administered two times)+Eth. Increasing the frequency of ionomycin stimulation from two (2-Ion+Eth) to three times (3-Ion+Eth) significantly (p<0.05) increased the cell number per embryo at the blastocyst stage. In addition, spermatozoa were pretreated with dithiothreitol (DTT), glutathione (GSH) or GSH+lysolecithin (LL) and used for producing bovine ICSI embryos. The blastocyst rate of bovine ICSI embryos produced from sperm pretreated with GSH was significantly (p<0.05) higher than those of embryos produced from sperm pretreated with DTT and GSH+LL. In conclusion, the embryo activation methods and sperm pretreatments examined in the present study did not affect the normal fertilization rate of bovine ICSI embryos. However, activation with 3-Ion+Eth and sperm pretreatment with GSH increased the cell number per embryo at blastocyst stage and the blastocyst rate, respectively, in bovine ICSI embryos.
authors.affiliation: Department of Tropical Agriculture and International Cooperation, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Jai-Wei
authors.initials: JW
authors.lastname: Lee
authors.affiliation: Department of Animal Science, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Hsun-Chung
authors.initials: HC
authors.lastname: Chang
authors.affiliation: Department of Veterinary Medicine, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Hung-Yi
authors.initials: HY
authors.lastname: Wu
authors.affiliation: Department of Veterinary Medicine, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Shyh-Shyan
authors.initials: SS
authors.lastname: Liu
authors.affiliation: Livestock Research Institute, Council of Agriculture, Executive Yuan, Hsinhua, Taiwan.
authors.forename: Chih-Hua
authors.initials: CH
authors.lastname: Wang
authors.affiliation: Graduate Institute of Animal Vaccine Technology, National Pingtung University of Science and Technology, Neipu, Taiwan.
authors.forename: Chun-Yen
authors.initials: CY
authors.lastname: Chu
authors.affiliation: Department of Animal Science, National Pingtung University of Science and Technology, Neipu, Taiwan. Electronic address: pcshen@mail.npust.edu.tw.
authors.forename: Perng-Chih
authors.initials: PC
authors.lastname: Shen
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D004229:Dithiothreitol / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D005260:Female; D005978:Glutathione / Q000494:pharmacology; D015759:Ionomycin / Q000494:pharmacology; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008297:Male; D020554:Sperm Injections, Intracytoplasmic / Q000379:methods*; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*
chemical_list: D008244:Lysophosphatidylcholines; D015759:Ionomycin; D005978:Glutathione; D004229:Dithiothreitol
keywords: Activation method; Bovine; Intracytoplasmic sperm injection; Sperm pretreatment