A zebrafish model of tauopathy allows in vivo imaging of neuronal cell death and drug evaluation
Dominik Paquet, … , Bettina Schmid, Christian Haass
Dominik Paquet, … , Bettina Schmid, Christian Haass
Published April 13, 2009
Citation Information: J Clin Invest. 2009;119(5):1382-1395. https://doi.org/10.1172/JCI37537.
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Technical Advance Neuroscience

A zebrafish model of tauopathy allows in vivo imaging of neuronal cell death and drug evaluation

Abstract

Our aging society is confronted with a dramatic increase of patients suffering from tauopathies, which include Alzheimer disease and certain frontotemporal dementias. These disorders are characterized by typical neuropathological lesions including hyperphosphorylation and subsequent aggregation of TAU protein and neuronal cell death. Currently, no mechanism-based cures are available. We generated fluorescently labeled TAU transgenic zebrafish, which rapidly recapitulated key pathological features of tauopathies, including phosphorylation and conformational changes of human TAU protein, tangle formation, neuronal and behavioral disturbances, and cell death. Due to their optical transparency and small size, zebrafish larvae are well suited for both in vivo imaging and drug development. TAU-induced neuronal cell death was imaged by time-lapse microscopy in vivo. Furthermore, we used this zebrafish model to identify compounds targeting the TAU kinase glycogen synthase kinase 3β (GSK3β). We identified a newly developed highly active GSK3β inhibitor, AR-534, by rational drug design. AR-534 reduced TAU phosphorylation in TAU transgenic zebrafish. This transgenic zebrafish model may become a valuable tool for further studies of the neuropathology of dementia.

Authors

Dominik Paquet, Ratan Bhat, Astrid Sydow, Eva-Maria Mandelkow, Stefan Berg, Sven Hellberg, Johanna Fälting, Martin Distel, Reinhard W. Köster, Bettina Schmid, Christian Haass

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Figure 1

A Gal4/UAS–based bidirectional expression system in zebrafish.

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A Gal4/UAS–based bidirectional expression system in zebrafish. (A) The D...
(A) The Driver construct contains the neuronal zebrafish promoter HuC driving the expression of Gal4-VP16, which binds to the UAS on the Responder construct. Here, it activates the bidirectional expression of hTAU-P301L and DsRed via the minimal promoters. UAS-dependent gene expression of TAU and DsRed is indicated in living fish by DsRed fluorescence. Driver and Responder constructs are flanked by Tol2 transposon sites. (B) To generate transgenic fish, the Driver and Responder constructs were mixed and injected together with Tol2 mRNA. The mRNA is translated to active transposase, which detects the flanking Tol2 elements and catalyzes random integration into the zebrafish genome in a subset of embryonic cells for a short time period, generating mosaic founder embryos. Mosaic DsRed-positive larvae were raised and outcrossed with wild-type fish. A subset of the offspring will be transgenic and can be easily identified and sorted by DsRed-positive neurons. Scale bar: 1 mm. (C) Double immunostainings for total TAU (T46 antibody) and DsRed of 32-hpf transgenic zebrafish embryos expressing hTAU-P301L and DsRed. Transgenic embryos express both hTAU-P301L and DsRed in spinal cord neurons, showing effective bidirectional expression from the Responder construct. Lateral views of the trunk above the end of the yolk extension, anterior to the left. Scale bar: 20 μm.