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ZFIN ID: ZDB-PUB-161013-5
Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons
Weber, T., Namikawa, K., Winter, B., Müller-Brown, K., Kühn, R., Wurst, W., Köster, R.W.
Date: 2016
Source: Development (Cambridge, England)   143(22): 4279-4287 (Journal)
Registered Authors: Köster, Reinhard W., Namikawa, Kazuhiko, Weber, Thomas
Keywords: Zebrafish, Cerebellum, Purkinje cells, Cell ablation, ApoptosisInducible caspase
MeSH Terms:
  • Animals
  • Animals, Genetically Modified
  • Apoptosis/drug effects
  • Apoptosis/genetics*
  • Caspase 8/genetics*
  • Cell Death/drug effects
  • Cell Death/genetics
  • Cell Survival/genetics
  • Cerebellum/cytology*
  • Cerebellum/drug effects
  • Cerebellum/metabolism
  • Genes, Reporter/drug effects
  • Genes, Transgenic, Suicide*/drug effects
  • Phagocytosis/genetics
  • Purkinje Cells/drug effects
  • Purkinje Cells/physiology*
  • Tamoxifen/pharmacology
  • Zebrafish*/embryology
  • Zebrafish*/genetics
PubMed: 27729409 Full text @ Development
FIGURES
ABSTRACT
The zebrafish is a well-established model organism in which to study in vivo mechanisms of cell communication, differentiation and function. Existing cell ablation methods are either invasive or they rely on the cellular expression of prokaryotic enzymes and the use of antibiotic drugs as cell death-inducing compounds. We have recently established a novel inducible genetic cell ablation system based on tamoxifen-inducible Caspase 8 activity, thereby exploiting mechanisms of cell death intrinsic to most cell types. Here, we prove its suitability in vivo by monitoring the ablation of cerebellar Purkinje cells (PCs) in transgenic zebrafish that co-express the inducible caspase and a fluorescent reporter. Incubation of larvae in tamoxifen for 8 h activated endogenous Caspase 3 and cell death, whereas incubation for 16 h led to the near-complete loss of PCs by apoptosis. We observed synchronous cell death autonomous to the PC population and phagocytosing microglia in the cerebellum, reminiscent of developmental apoptosis in the forebrain. Thus, induction of apoptosis through targeted activation of caspase by tamoxifen (ATTACTM) further expands the repertoire of genetic tools for conditional interrogation of cellular functions.
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