Wang Jinliang, Williams Christopher K, DeTure Michael A, Magaki Shino D, Dickson Dennis W, Vinters Harry V, Seidler Paul M
Department of Pharmacology and Pharmaceutical Sciences, University of Southern California Mann School of Pharmacy and Pharmaceutical Sciences, 1985 Zonal Avenue, Los Angeles, CA 90089-9121, USA.
Department of Pathology and Laboratory Medicine, David Geffen School of Medicine at University of California Los Angeles, Los Angeles, CA 90095, USA.
Structure. 2024 Dec 5;32(12):2251-2258.e3. doi: 10.1016/j.str.2024.10.013. Epub 2024 Nov 1.
Fibril-type aggregates of tau occur in Alzheimer's disease (AD) and dozens of tauopathies. Fibrils catalyze aggregation by prion-like seeding, which in part underlies disease progression. Seeding by recombinant and brain-derived tau fibrils is measured using biosensor cells that express aggregation-prone tau mutants fused with fluorescent reporter proteins. Seeding results in a punctated phenotype that is well established, but evidence that fluorescent tau fusion proteins from biosensor cells assemble into fibril-type structures is lacking. We investigated the effects of seeding on fibril formation by biosensor cells. Fluorescent punctated cell phenotypes that were catalyzed persisted with varying stabilities. Seeded cells bearing punctated phenotypes yielded sarkosyl-insoluble fibrils, although non-seeded cells did not. ImmunoEM of cell-purified fibrils shows that GFP localizes to the proteolytically sensitive fuzzy coat of tau fibrils. The presented data offer compelling evidence that fluorescent puncta are fibril-type aggregates of tau that result from prion-like seeding.
tau蛋白的原纤维型聚集体出现在阿尔茨海默病(AD)和数十种tau蛋白病中。原纤维通过朊病毒样种子催化聚集,这在一定程度上是疾病进展的基础。使用表达与荧光报告蛋白融合的易聚集tau突变体的生物传感器细胞来检测重组tau原纤维和脑源性tau原纤维的种子作用。种子作用会导致一种已被充分证实的点状表型,但缺乏生物传感器细胞中的荧光tau融合蛋白组装成原纤维型结构的证据。我们研究了种子作用对生物传感器细胞原纤维形成的影响。由种子作用催化产生的荧光点状细胞表型以不同的稳定性持续存在。具有点状表型的种子细胞产生了对 Sarkosyl不溶性原纤维,而非种子细胞则没有。对细胞纯化原纤维的免疫电镜显示,绿色荧光蛋白(GFP)定位于tau原纤维对蛋白酶敏感的模糊外层。所呈现的数据提供了令人信服的证据,即荧光点是由朊病毒样种子作用产生的tau蛋白原纤维型聚集体。