title: Colcemid treatment during oocyte maturation improves preimplantation development of cloned pig embryos by influencing meiotic progression and cytoplasmic maturation.
abstract: The objective of this study was to examine the effects of colcemid treatment during oocyte in vitro maturation (IVM) and embryonic development after parthenogenetic activation (PA) and somatic-cell nucleus transfer (SCNT) in pigs. Immature oocytes were treated with colcemid from 0 to 22, 38 to 42, or 0 to 22 hr followed by 38 to 42 hr during IVM (designated as COL0-22, COL38-42, and COL0-22/38-42, respectively). The proportion of oocytes reaching the germinal vesicle (GV)/GV breakdown (GVBD) stage after 22 hr of IVM was higher in COL0-22 (98.4%) than in controls not exposed to colcemid (68.7%). The proportion of metaphase-II (MII) oocytes after 30 hr of IVM was higher in control (79.6%) than in COL0-22 oocytes (61.7%); overall nuclear progression to the MII stage was not influenced by colcemid treatment by the end of the IVM period (93.8, 86.7, 86.8, and 84.8% for control, COL0-22, COL38-42, and COL0-22/38-42, respectively). COL0-22 oocytes showed higher intra-oocyte glutathione content (1.7 vs. 1.0-1.3 pixels/oocyte) and increased blastocyst formation after PA (68.7% vs. 42.5-52.2%) and SCNT (39.4% vs. 16.3-28.6%) than control, COL38-42, and COL0-22/38-42 oocytes. Colcemid treatment for 0-22 and 0-22/38-42 hr of IVM also stimulated the expression of cyclin-dependent kinase 1 (CDK1), proliferating cell nuclear antigen (PCNA), and extracellular signal-regulated kinase 2 (ERK2) mRNAs. Our results thus demonstrate that the presence of colcemid during the early stage of IVM stimulates preimplantation development of PA and SCNT porcine embryos by improving the cytoplasmic microenvironment.
authors.affiliation: College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.
authors.forename: Joohyeong
authors.initials: J
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Konkuk University, Seoul, Korea.
authors.forename: Jong-Im
authors.initials: JI
authors.lastname: Park
authors.affiliation: College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.|Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
authors.forename: Geun-Shik
authors.initials: GS
authors.lastname: Lee
authors.affiliation: College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.|Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
authors.forename: Jung Hoon
authors.initials: JH
authors.lastname: Choi
authors.affiliation: Division of Applied Animal Science, College of Animal Life Science, Kangwon National University, Chuncheon, Korea.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
authors.affiliation: Division of Applied Animal Science, College of Animal Life Science, Kangwon National University, Chuncheon, Korea.
authors.forename: Choon-Keun
authors.initials: CK
authors.lastname: Park
authors.affiliation: Department of Life Science, College of BioNano Technology, Gachon University, Incheon, Korea.
authors.forename: Dae Young
authors.initials: DY
authors.lastname: Kim
authors.affiliation: College of Veterinary Medicine, Chungbuk National University, Cheongju, Korea.
authors.forename: Sang-Hwan
authors.initials: SH
authors.lastname: Hyun
authors.affiliation: College of Veterinary Medicine, Kangwon National University, Chuncheon, Korea.|Institute of Veterinary Science, Kangwon National University, Chuncheon, Korea.
authors.forename: Eunsong
authors.initials: E
authors.lastname: Lee
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D019976:Cloning, Organism; D003593:Cytoplasm / Q000378:metabolism*; D003703:Demecolcine / Q000494:pharmacology*; D005260:Female; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects*; D008540:Meiosis / Q000187:drug effects*; D009865:Oocytes / Q000378:metabolism*; D013552:Swine
chemical_list: D003703:Demecolcine