title: Ploidy manipulation and induction of alternate cleavage patterns through inhibition of centrosome duplication in the early zebrafish embryo.
abstract: BACKGROUND
  Whole genome duplication is a useful genetic tool because it allows immediate and complete genetic homozygosity in gynogenetic offspring. A whole genome duplication method in zebrafish, Heat Shock, involves a heat pulse in the period 13-15 min postfertilization (mpf) to inhibit cytokinesis of the first mitotic cycle. However, Heat Shock produces a relatively low yield of gynogenotes.
  
  
  RESULTS
  A heat pulse at a later time point during the first cell cycle (22 mpf, HS2) results in a high (>80%) frequency of embryos exhibiting a precise one-cell division stall during the second cell cycle, inducing whole genome duplication. Coupled with haploid production, HS2 generates viable gynogenetic diploids with yields up to 4 times higher than those achieved through standard Heat Shock. The cell cycle delay also causes blastomere cleavage pattern variations, supporting a role for cytokinesis in spindle orientation during the following cell cycle.
  
  
  CONCLUSIONS
  Our studies provide a new tool for whole genome duplication, induced gynogenesis, and cleavage pattern alteration in zebrafish, based on a time period before the initiation of cell division that is sensitive to temperature-mediated interference with centrosome duplication. Targeting of this period may also facilitate genetic and developmental manipulations in other organisms.
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Madison, Wisconsin.
authors.forename: Jonathon
authors.initials: J
authors.lastname: Heier
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Madison, Wisconsin.
authors.forename: Kendra A
authors.initials: KA
authors.lastname: Takle
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Madison, Wisconsin.
authors.forename: Andrew O
authors.initials: AO
authors.lastname: Hasley
authors.affiliation: Laboratory of Genetics, University of Wisconsin - Madison, 425-G Henry Mall, Madison, Wisconsin.
authors.forename: Francisco
authors.initials: F
authors.lastname: Pelegri
mesh_terms: D000818:Animals; D002453:Cell Cycle; D018385:Centrosome / Q000502:physiology*; D002970:Cleavage Stage, Ovum; D004626:Embryology / Q000379:methods*; D005821:Genetic Techniques; D006720:Homozygote; D006358:Hot Temperature; D009154:Mutation; D011003:Ploidies; D015027:Zebrafish
keywords: centrioles; gynogenesis; heat shock; parthenogenesis; ploidy manipulation; tetraploidy; whole genome duplication; zebrafish