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Development. 2016 Nov 15;143(22):4279-4287. Epub 2016 Oct 11.

Caspase-mediated apoptosis induction in zebrafish cerebellar Purkinje neurons.

Author information

1
TU Braunschweig, Zoological Institute, Cellular and Molecular Neurobiology, Spielmannstr. 7, Braunschweig 38106, Germany.
2
Helmholtz Zentrum München, German Research Center for Environmental Health, Institute of Developmental Genetics, Ingolstädter Landstr. 1, Neuherberg 85764, Germany.
3
Deutsches Zentrum für Neurodegenerative Erkrankungen e. V. (DZNE), Standort München, Feodor-Lynen-Str. 17, München 81377, Germany.
4
Munich Cluster for Systems Neurology (SyNergy), Feodor-Lynen-Str. 17, München 81377, Germany.
5
Technische Universität München-Weihenstephan, Lehrstuhl für Entwicklungsgenetik, c/o Helmholtz Zentrum München, Ingolstädter Landstr. 1, Neuherberg 85764, Germany.
6
TU Braunschweig, Zoological Institute, Cellular and Molecular Neurobiology, Spielmannstr. 7, Braunschweig 38106, Germany r.koester@tu-bs.de.

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.

KEYWORDS:

Apoptosis; Cell ablation; Cerebellum; Inducible caspase; Purkinje cells; Zebrafish

PMID:
27729409
DOI:
10.1242/dev.122721
[Indexed for MEDLINE]
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