Tau aggregation and toxicity in a cell culture model of tauopathy.
Intracellular aggregation of the microtubule-associated protein tau into filamentous inclusions is a defining characteristic of Alzheimer disease. Because appearance of tau-aggregate bearing lesions correlates with both cognitive decline and neurodegeneration, it has been hypothesized that tau aggregation may be directly toxic to cells that harbor them. Testing this hypothesis in cell culture has been complicated by the resistance of full-length tau isoforms to aggregation over experimentally tractable time periods. To overcome this limitation, a small-molecule agonist of the tau aggregation reaction, Congo red, was used to drive aggregation within HEK-293 cells expressing full-length tau isoform htau40. Formation of detergent-insoluble aggregates was both time and agonist concentration dependent. At 10 microM Congo red, detergent-insoluble aggregates appeared with pseudo-first order kinetics and a half-life of approximately 5 days. By 7 days in culture, total tau levels increased 2-fold, with approximately 30% of total tau converted into detergent-insoluble aggregates. Agonist addition also led to rapid losses in the tubulin binding activity of tau, although tau was not hyperphosphorylated as judged by occupancy of phosphorylation sites Ser396/Ser404. Tau aggregation was associated with decreased viability as detected by ToPro-3 uptake. The results, which establish a new approach for analysis of tau aggregation in cells independent of tau hyperphosphorylation, suggest that conformational changes associated with aggregation are incompatible with microtubule binding, and that toxicity associated with intracellular tau aggregation is not acute but develops over a period of days.
Bandyopadhyay
Bhaswati
B
Center for Molecular Neurobiology, the Department of Molecular and Cellular Biochemistry, Ohio State University, Columbus 43210, USA.
Li
Guibin
G
Yin
Haishan
H
Kuret
Jeff
J
D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D004396:Coloring Agents / Q000494:pharmacology*; D003224:Congo Red / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D008565:Membrane Proteins / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D008954:Models, Biological; D010766:Phosphorylation / Q000187:drug effects; D011485:Protein Binding / Q000187:drug effects; D020033:Protein Isoforms / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000187:drug effects*; D013997:Time Factors; D014404:Tubulin / Q000378:metabolism; D016875:tau Proteins
D004396:Coloring Agents; D003224:Congo Red; D008565:Membrane Proteins; D020033:Protein Isoforms; D014404:Tubulin; C054369:MAPT protein, human; D016875:tau Proteins