title: The effects of demecolcine, alone or in combination with sucrose on bovine oocyte protrusion rate, MAPK1 protein level and c-mos gene expression level.
abstract: OBJECTIVE
  Our study aims to clarify the effects of demecolcine, alone or in combination with sucrose on bovine oocyte protrusion rate, MAPK1 protein level and c-mos gene expression level.
  
  
  METHODS
  The effects of the demecolcine concentration, treatment duration, and synergistic effects with sucrose solution on the rate of membrane protrusions of bovine oocytes were investigated. Using real-time fluorescent quantitative PCR, western blot analysis, and immunofluorescence assays, the expression of the maternal c-mos gene, the protein level of mitogen-activated protein kinase 1 (MAPK1), and the change in the localization of spindles and nuclei during the demecolcine treatment were analyzed in bovine oocytes.
  
  
  RESULTS
  Treatment of bovine oocytes with both demecolcine (0.6 μg/mL) and sucrose (0.05 M) for 1 h led to the highest rate of membrane protrusions, and synergistic effects were also observed. Real-time fluorescent quantitative PCR analysis revealed that the demecolcine treatment up-regulated the expression of the maternal c-mos gene. Western blot analysis indicated that the demecolcine treatment enhanced the protein level of MAPK1 in bovine oocytes. Immunofluorescence analysis indicated that the spindles and nuclei were localized at the place of the membrane protrusions.
  
  
  CONCLUSIONS
  The present results suggest that demecolcine might contribute to the activation of the Mos/MAPK pathway and affect spindle structure. These results provide a reference for more efficient generation of enucleated bovine oocytes.
authors.affiliation: Laboratory Animal Center, College of Animal Sciences, Jilin University, Changchun, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Gao
authors.forename: Jiusheng
authors.initials: J
authors.lastname: Ren
authors.forename: Liqun
authors.initials: L
authors.lastname: Zhang
authors.forename: Yonghong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xinhui
authors.initials: X
authors.lastname: Wu
authors.forename: Hao
authors.initials: H
authors.lastname: Jiang
authors.forename: Fangfang
authors.initials: F
authors.lastname: Xu
authors.forename: Bao
authors.initials: B
authors.lastname: Yuan
authors.forename: Xianfeng
authors.initials: X
authors.lastname: Yu
authors.forename: Jiabao
authors.initials: J
authors.lastname: Zhang
mesh_terms: D000818:Animals; D002417:Cattle; D002467:Cell Nucleus / Q000187:drug effects; D003703:Demecolcine / Q000008:administration & dosage*; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000096:biosynthesis*; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D017231:Proto-Oncogene Proteins c-mos / Q000096:biosynthesis*; D008941:Spindle Apparatus / Q000187:drug effects; D013395:Sucrose / Q000008:administration & dosage
chemical_list: D013395:Sucrose; D017231:Proto-Oncogene Proteins c-mos; D019950:Mitogen-Activated Protein Kinase 1; D003703:Demecolcine