PUBLICATION

Ploidy manipulation and induction of alternate cleavage patterns through inhibition of centrosome duplication in the early zebrafish embryo

Authors
Heier, J., Takle, K.A., Hasley, A.O., Pelegri, F.
ID
ZDB-PUB-150808-7
Date
2015
Source
Developmental dynamics : an official publication of the American Association of Anatomists   244(10): 1300-12 (Journal)
Registered Authors
Pelegri, Francisco
Keywords
zebrafish, heat shock, centrioles, whole genome duplication, ploidy manipulation, tetraploidy, parthenogenesis, gynogenesis
MeSH Terms
  • Animals
  • Cell Cycle
  • Centrosome/physiology*
  • Cleavage Stage, Ovum*
  • Embryology/methods*
  • Genetic Techniques*
  • Homozygote
  • Hot Temperature
  • Mutation
  • Ploidies*
  • Zebrafish
PubMed
26249657 Full text @ Dev. Dyn.
Abstract
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 minutes post-fertilization (mpf) to inhibit cytokinesis of the first mitotic cycle. However, Heat Shock produces a relatively low yield of gynogenotes.
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.
Our studies provide a new tool for whole genome duplication, induced gynogenesis and cleavage pattern alteration in zebrafish, based on a time period prior to 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. This article is protected by copyright. All rights reserved.
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Phenotype
Mutation and Transgenics
Human Disease / Model Data
Sequence Targeting Reagents
Fish
Antibodies
Orthology
Engineered Foreign Genes
Mapping
Errata and Notes