Comparative Analysis of an Improved Thioflavin-S Stain, Gallyas Silver Stain, and Immunohistochemistry for Neurofibrillary Tangle Demonstration on the Same Sections

Sun, Anyang; Nguyen, Xuan V.; Bing, Guoying

doi:10.1177/002215540205000403

Cited by 124 publications

(94 citation statements)

References 29 publications

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“…Cells were then washed extensively in PBS before being incubated in a thioflavin-S solution (0.005%) (Sigma) for 10 min. The cells were then washed 3 times in 70% ethanol and once in water before mounting (33). Cells were viewed with a Nikon Diaphot 200 epifluorescence microscope, and images were captured with a Digital spot camera (Diagnostic Instruments), digitally stored, and displayed using the accompanying software.…”

Section: Methodsmentioning

confidence: 99%

Glycogen Synthase Kinase 3β Induces Caspase-cleaved Tau Aggregation in Situ

Cho¹,

Johnson

2004

Journal of Biological Chemistry

110

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Tau is a substrate of caspases, and caspase-cleaved tau has been detected in Alzheimer's disease brain but not in control brain. Furthermore, in vitro studies have revealed that caspase-cleaved tau is more fibrillogenic than full-length tau. Considering these previous findings, the purpose of this study was to determine how the caspase cleavage of tau affected tau function and aggregation in a cell model system. The effects of glycogen synthase kinase 3␤ (GSK3␤), a well established tau kinase, on these processes also were examined. Tau or tau that had been truncated at Asp-421 to mimic caspase cleavage (Tau-D421) was transfected into cells with or without GSK3␤, and phosphorylation, microtubule binding, and tau aggregation were examined. Tau-D421 was not as efficiently phosphorylated by GSK3␤ as fulllength tau. Tau-D421 efficiently bound microtubules, and in contrast to the full-length tau, co-expression with GSK3␤ did not result in a reduction in the ability of Tau-D421 to bind microtubules. In the absence of GSK3␤, neither Tau-D421 nor full-length tau formed Sarkosyl-insoluble inclusions. However, in the presence of GSK3␤, Tau-D421, but not full-length tau, was present in the Sarkosyl-insoluble fraction and formed thioflavin-Spositive inclusions in the cell. Nonetheless, co-expression of GSK3␤ and Tau-D421 did not result in an enhancement of cell death. These data suggest that a combination of phosphorylation events and caspase activation contribute to the tau oligomerization process in Alzheimer's disease, with GSK3␤-mediated tau phosphorylation preceding caspase cleavage.

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Section: Methodsmentioning

confidence: 99%

Glycogen Synthase Kinase 3β Induces Caspase-cleaved Tau Aggregation in Situ

Cho¹,

Johnson

2004

Journal of Biological Chemistry

110

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“…After removing the paraffin and rehydration, the autofluorescence was quenched (Sun et al, 2002), sections were incubated in 0.05% Thioflavin S (Sigma) for 8 min, and excess Thioflavin S was removed by two brief washes in 80% EtOH. After three washing steps in large volumes of tap water, the sections were mounted in Permafluor (Beckman Coulter, Paris, France).…”

Section: Histologymentioning

confidence: 99%

The Potential for β-Structure in the Repeat Domain of Tau Protein Determines Aggregation, Synaptic Decay, Neuronal Loss, and Coassembly with Endogenous Tau in Inducible Mouse Models of Tauopathy

Mocanu¹,

Nissen²,

Eckermann³

et al. 2008

J. Neurosci.

274

245

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We describe two new transgenic mouse lines for studying pathological changes of Tau protein related to Alzheimer's disease. They are based on the regulatable expression of the four-repeat domain of human Tau carrying the FTDP17 (frontotemporal dementia and parkinsonism linked to chromosome 17) mutation ⌬K280 (Tau RD /⌬K280), or the ⌬K280 plus two proline mutations in the hexapeptide motifs (Tau RD /⌬K280/I277P/I308P). The ⌬K280 mutation accelerates aggregation ("proaggregation mutant"), whereas the proline mutations inhibit Tau aggregation in vitro and in cell models ("antiaggregation mutant"). The inducible transgene expression was driven by the forebrain-specific CaMKII␣ (calcium/calmodulin-dependent protein kinase II␣) promoter. The proaggregation mutant leads to Tau aggregates and tangles as early as 2-3 months after gene expression, even at low expression (70% of endogenous mouse Tau). The antiaggregation mutant does not aggregate even after 22 months of gene expression. Both mutants show missorting of Tau in the somatodendritic compartment and hyperphosphorylation in the repeat domain [KXGS motifs, targets of the kinase MARK (microtubule affinity regulating kinase)]. This indicates that these changes are related to Tau expression rather than aggregation. The proaggregation mutant causes astrogliosis, loss of synapses and neurons from 5 months of gene expression onward, arguing that Tau toxicity is related to aggregation. Remarkably, the human proaggregation mutant Tau RD coaggregates with mouse Tau, coupled with missorting and hyperphosphorylation at multiple sites. When expression of proaggregation Tau RD is switched off, soluble and aggregated exogenous Tau RD disappears within 1.5 months. However, tangles of mouse Tau, hyperphosphorylation, and missorting remain, suggesting an extended lifetime of aggregated wild-type Tau once a pathological conformation and aggregation is induced by a proaggregation Tau species.

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“…Cells were then washed extensively in PBS before being incubated in a thioflavin-S solution (Sigma) (0.005%) for 10 min. The cells were then washed in 70% ethanol and water before mounting [17]. Fig.…”

Section: Immunocytochemistrymentioning

confidence: 99%