title: Effects of activation methods and culture conditions on development of parthenogenetic porcine embryos.
abstract: The effects of different activation methods and culture conditions on early development of porcine parthenotes were examined. Three different activation methods were tested: (1) electroporation; (2) electroporation followed by incubation in the presence of butyrolactone I, an inhibitor of cdc2 and cdk2 kinases; and (3) electroporation followed by a treatment with cycloheximide, a blocker of protein synthesis. The activated oocytes were cultured in two different media, NCSU-23 and PZM-3 under 5% CO2 in air. In a separate experiment, the effects of high (approximately 20%) or low (5%) O2 tension on early embryo development were also evaluated. The average pronuclear formation was less (p<0.05) in the electroporated oocytes (83.9+/-1.7%) compared with those activated by electroporation and butyrolactone I or electroporation plus cycloheximide (92.8+/-0.8 and 93.0+/-1.0%). In PZM-3 medium, the average frequencies of blastocyst formation (59.7+/-3.6%) and hatching (10.6+/-1.3%) were greater than those in NCSU-23 medium (39.9+/-3.1% blastocyst formation, p<0.05; and 0.2+/-0.2% hatching; p<0.001). Furthermore, the average nuclear number was also greater (p<0.001) in blastocysts developed in PZM-3 (50.2+/-1.3) than in those developed in NCSU-23 (35.3+/-1.1). Blastocyst formation was similar (p>0.10) among the three activation procedures when parthenotes were cultured in NCSU-23, while in PZM-3 more (p<0.05) parthenotes produced by electroporation plus butyrolactone or electroporation plus cycloheximide developed into blastocysts compared to electroporation alone (64.9+/-5.2 and 68.6+/-3.5% compared with 45.6+/-4.7%). Incidences of apoptotic nuclei were similar (p>0.10) among all treatments. No difference in development was found between parthenotes that developed under high versus low O2 tension (p>0.10). These results demonstrate that activation methods targeting the calcium signaling pathway at several points trigger embryonic development more efficiently than electroporation alone. The data also imply that the PZM-3 medium provides for enhanced culture conditions for the early development of parthenogenetic porcine embryos than NCSU-23.
authors.affiliation: Department of Animal Breeding Science, Center of Agricultural Sciences, University of Debrecen, Böszörményi Street 138, Debrecen, Hungary.
authors.forename: László
authors.initials: L
authors.lastname: Nánássy
authors.forename: Kiho
authors.initials: K
authors.lastname: Lee
authors.forename: András
authors.initials: A
authors.lastname: Jávor
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Macháty
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods / Q000662:veterinary; D003470:Culture Media; D003513:Cycloheximide / Q000494:pharmacology*; D018274:Electroporation / Q000662:veterinary*; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D005260:Female; D009865:Oocytes / Q000502:physiology*; D010100:Oxygen / Q000494:pharmacology; D010312:Parthenogenesis / Q000187:drug effects / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013552:Swine / Q000196:embryology* / Q000502:physiology
chemical_list: D003470:Culture Media; D047428:Protein Kinase Inhibitors; D011500:Protein Synthesis Inhibitors; C082868:butyrolactone I; D003513:Cycloheximide; D015107:4-Butyrolactone; D010100:Oxygen