Tau-proximity ligation assay reveals extensive previously undetected pathology prior to neurofibrillary tangles in preclinical Alzheimer's disease.
Tau-proximity ligation assay reveals extensive previously undetected pathology prior to neurofibrillary tangles in preclinical Alzheimer's disease.
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DOI：
10.1186/s40478-020-01117-y
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发表时间：
2021-01-28
影响因子：
7.1
通讯作者：
Alegre-Abarrategui J
中科院分区：
文献类型：
--
作者：
Bengoa-Vergniory N;Velentza-Almpani E;Silva AM;Scott C;Vargas-Caballero M;Sastre M;Wade-Martins R;Alegre-Abarrategui J
多聚化是蛋白质样疾病，如阿尔茨海默病(AD)的关键过程，因为它是自我模板化tau和β-淀粉样淀粉样蛋白发生所必需的。AT8-过度磷酸化tau的免疫组织化学目前用于tau病理的诊断和分期。鉴于tau-tau的相互作用可以在没有过度磷酸化或其他翻译后修饰(PTM)的情况下发生，直接观察tau的多聚体可以发现早期的病理性tau多聚体。在这里，我们使用双分子荧光互补，依赖于雷帕霉素的FKBP/FRB-tau相互作用和透射电子显微镜来证明tau-邻近连接试验(tau-pla)的体外特异性。然后，我们用tau-PLA分析了MAPT KO和P301S转基因小鼠，以及所有Braak阶段的人海马体和颞叶等皮质，并将其与诊断抗体AT8、早期磷酸化依赖的AT180和构象依赖的抗体MC1的免疫组织化学进行了比较。最后，我们进行了蛋白酶-K处理，以推测淀粉样β折叠的含量。我们新的tau-邻近连接实验(tau-pla)直接在原位观察了tau-tau的相互作用，并且只能识别tau多聚体而不能识别单体。在MAPT KO小鼠脑内未发现信号，但被广泛标记为P301S转基因小鼠和AD脑。发现了两组结构，一种是以前未见报道的广泛的小规模弥漫性病变，另一组是大型的神经原纤维样病变。Tau-pla标记的弥漫性病变最早出现在Braak分期，大多不伴有缠结状tau免疫组织化学，在大多数区域0-II期定量的AT180-、AT8-和MC1-免疫组织化学标记的小点状/线状病理明显更敏感。与大的病变相比，tau-pla标记的弥漫性病变对蛋白水解酶-K极为敏感。Tau-pla是第一种在体外和原位直接显示tau多聚体的方法，具有很高的特异性。我们发现，tau多聚化在症状前Braak早期广泛出现，是一种以前未见报道的弥漫性病理类型。重要的是，在我们的研究中，多聚化是AD tau病理中最早可检测到的分子事件。我们的发现为早期tau病理的研究打开了新的窗口，对早期诊断和治疗策略的设计具有潜在的意义。本文的在线版本(10.1186/s40478-020-020-y)包含向授权用户提供的补充材料。
Multimerization is a key process in prion-like disorders such as Alzheimer’s disease (AD), since it is a requirement for self-templating tau and beta-amyloid amyloidogenesis. AT8-immunohistochemistry for hyperphosphorylated tau is currently used for the diagnosis and staging of tau pathology. Given that tau–tau interactions can occur in the absence of hyperphosphorylation or other post-translational modifications (PTMs), the direct visualization of tau multimerization could uncover early pathological tau multimers. Here, we used bimolecular fluorescent complementation, rapamycin-dependent FKBP/FRB-tau interaction and transmission electron microscopy to prove the in vitro specificity of tau-proximity ligation assay (tau-PLA). We then analyzed MAPT KO and P301S transgenic mice, and human hippocampus and temporal isocortex of all Braak stages with tau-PLA and compared it with immunohistochemistry for the diagnostic antibody AT8, the early phosphorylation-dependent AT180, and the conformational-dependent antibody MC1. Finally, we performed proteinase-K treatment to infer the content of amyloidogenic beta-sheet fold. Our novel tau-proximity ligation assay (tau-PLA) directly visualized tau–tau interactions in situ, and exclusively recognized tau multimers but not monomers. It elicited no signal in MAPT KO mouse brains, but extensively labelled P301S transgenic mice and AD brain. Two groups of structures were detected, a previously unreported widespread small-sized diffuse pathology and large, neurofibrillary-like lesions. Tau-PLA-labelled diffuse pathology appeared from the earliest Braak stages, mostly unaccompanied by tangle-like tau-immunohistochemistry, being significantly more sensitive than any small-sized dot-/thread-like pathology labelled by AT180-, AT8- and MC1-immunohistochemistry in most regions quantified at stages 0-II. Tau-PLA-labelled diffuse pathology was extremely sensitive to Proteinase-K, in contrast to large lesions. Tau-PLA is the first method to directly visualize tau multimers both in vitro and in situ with high specificity. We find that tau multimerization appears extensively from the earliest presymptomatic Braak stages as a previously unreported type of diffuse pathology. Importantly, in our study multimerization is the earliest detectable molecular event of AD tau pathology. Our findings open a new window to the study of early tau pathology, with potential implications in early diagnosis and the design of therapeutic strategies. The online version of this article (10.1186/s40478-020-01117-y) contains supplementary material, which is available to authorized users.
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DOI：
10.1074/jbc.m114.589309
发表时间：
2015-01-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Falcon B;Cavallini A;Angers R;Glover S;Murray TK;Barnham L;Jackson S;O'Neill MJ;Isaacs AM;Hutton ML;Szekeres PG;Goedert M;Bose S
通讯作者：
Bose S
影响因子：
1.2
作者：
Furman, Jennifer L.;Holmes, Brandon B.;Diamond, Marc I.
通讯作者：
Diamond, Marc I.
影响因子：
16.2
作者：
GOEDERT, M;SPILLANTINI, MG;CROWTHER, RA
通讯作者：
CROWTHER, RA
影响因子：
9.9
作者：
ARRIAGADA, PV;GROWDON, JH;HYMAN, BT
通讯作者：
HYMAN, BT
影响因子：
16.2
作者：
de Calignon A;Polydoro M;Suárez-Calvet M;William C;Adamowicz DH;Kopeikina KJ;Pitstick R;Sahara N;Ashe KH;Carlson GA;Spires-Jones TL;Hyman BT
通讯作者：
Hyman BT