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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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 6

Chemical structure, design, and characteristics of GSK3 inhibitors AR-164 and AR-534.

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Chemical structure, design, and characteristics of GSK3 inhibitors AR-16...
(A) Left panel shows surface representation of the x-ray structure of GSK3β with inhibitor AR-164 in the active site. Polar areas are colored blue and lipophilic areas red. The methylpiperazine sulfone amide extends out toward the solvent area. Right panel shows top view of the active site of GSK3β. Dotted yellow lines represent hydrogen bonds between the protein backbone and AR-164. The 6-membered aromatic pyrazine moiety binds together with its anilino function to the backbone of the kinase. The pyridine ring binds to the conserved salt bridge formed by Lys85 and Asp200 (x-ray resolution: 2.47 υ). AR-534 binds in a similar way (not shown). (B and C) Chemical structure of AR-164 and AR-534 and compound characterization values including Ki (mean from 3 independent experiments performed in duplicate), selectivity over CDK2, solubility and permeability coefficient Pe (cm/min) over cell membranes (Caco2 cells), and BBB. (D) AR-164 and AR-534 inhibit pSer396 TAU phosphorylation in 3T3 fibroblasts harvested at 4 hours after treatment in comparison with total TAU, as analyzed by quantitative WB. (E) Graphical representation with IC50 values of AR-164 and AR-534 effects on inhibition of TAU phosphorylation compared with SB-415286 and LiCl. (F) Alignment of protein sequences of human and zebrafish GSK3β. Over 90% of the amino acids are identical; the residues in the active site of the enzyme, which interact with the inhibitors, are completely conserved.