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 2

Expression of hTAU-P301L induces rapid pathological hyperphosphorylation, conformational changes, and aggregation of TAU in transgenic zebrafish.

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Expression of hTAU-P301L induces rapid pathological hyperphosphorylation...
(A) Double whole-mount immunostainings for phosphorylated and total TAU of 32-hour-old transgenic zebrafish embryos expressing hTAU-P301L and DsRed. TAU is phosphorylated in spinal cord neurons at residues Thr231/Ser235 (AT180), Thr181 (AT270), Ser262/Ser356 (12E8), Ser396/Ser404 (PHF1), Ser422 (422), and Ser202/Thr205 (AT8). (B) WBs of total and phosphorylated TAU of 48-hour-old transgenic zebrafish embryos expressing hTAU-P301L and DsRed or DsRed alone or nontransgenic siblings. Phosphorylated TAU is only detected in TAU/DsRed transgenic embryos. No cross-reacting bands are detectable in controls at the same molecular weight. In addition to the specific band above 64 kDa (arrow) in TAU-positive embryos, antibodies PHF1 and 422 detect a nonspecific band at lower molecular weight in all embryos (asterisk). (C) TAU changes its conformation to a pathologic state, as shown in whole-mount immunostainings, by using the conformation-specific antibody MC1, in most neurons of 32-hour-old embryos expressing the TAU transgene. (D) Side views of 5-week-old living zebrafish in bright field and DsRed illumination. The fish are still rather translucent, and the transgene-expressing cells can be detected by their red fluorescence (arrowheads). (E) Immunohistochemical stainings of spinal cord paraffin sections of the same 5-week-old TAU transgenic zebrafish for total TAU (antibody DA9) and pathologically phosphorylated TAU (AT8) (arrowheads). (F) In addition, tangles are observed by Gallyas silver staining in sections of the same 5-week-old TAU transgenic zebrafish (arrowheads). Scale bars: 20 μm.