Laboratory of Psychiatry and Experimental Alzheimer's Research, Medical University of Innsbruck, 6020 Innsbruck, Austria.
Biomolecules. 2024 Jan 30;14(2):165. doi: 10.3390/biom14020165.
Alzheimer's disease (AD) is a progressive neurodegenerative disorder that is characterized by amyloid-beta (Aβ) plaques and tau neurofibrillary tangles (NFT). Modelling aspects of AD is challenging due to its complex multifactorial etiology and pathology. The present study aims to establish a cost-effective and rapid method to model the two primary pathologies in organotypic brain slices. Coronal hippocampal brain slices (150 µm) were generated from postnatal (day 8-10) C57BL6 wild-type mice and cultured for 9 weeks. Collagen hydrogels containing either an empty load or a mixture of human Aβ42 and P301S aggregated tau were applied to the slices. The media was further supplemented with various intracellular pathway modulators or heavy metals to augment the appearance of Aβ plaques and tau NFTs, as assessed by immunohistochemistry. Immunoreactivity for Aβ and tau was significantly increased in the ventral areas in slices with a mixture of human Aβ42 and P301S aggregated tau compared to slices with empty hydrogels. Aβ plaque- and tau NFT-like pathologies could be induced independently in slices. Heavy metals (aluminum, lead, cadmium) potently augmented Aβ plaque-like pathology, which developed intracellularly prior to cell death. Intracellular pathway modulators (scopolamine, wortmannin, MHY1485) significantly boosted tau NFT-like pathologies. A combination of nanomolar concentrations of scopolamine, wortmannin, MHY1485, lead, and cadmium in the media strongly increased Aβ plaque- and tau NFT-like immunoreactivity in ventral areas compared to the slices with non-supplemented media. The results highlight that we could harness the potential of the collagen hydrogel-based spreading of human Aβ42 and P301S aggregated tau, along with pharmacological manipulation, to produce pathologies relevant to AD. The results offer a novel ex vivo organotypic slice model to investigate AD pathologies with potential applications for screening drugs or therapies in the future.
阿尔茨海默病（AD）是一种进行性神经退行性疾病，其特征是淀粉样β（Aβ）斑块和 tau 神经原纤维缠结（NFT）。由于其复杂的多因素病因和病理学，模拟 AD 的各个方面具有挑战性。本研究旨在建立一种经济高效且快速的方法，在器官型脑片中模拟两种主要的病理学。从出生后（第 8-10 天）的 C57BL6 野生型小鼠中生成冠状海马脑片（150 µm），并培养 9 周。将含有空载体或人 Aβ42 和 P301S 聚集 tau 的混合物的胶原水凝胶施加到切片上。通过免疫组织化学进一步用各种细胞内途径调节剂或重金属补充培养基，以增加 Aβ 斑块和 tau NFT 的出现。与含有空水凝胶的切片相比，含有人 Aβ42 和 P301S 聚集 tau 的混合物的切片中，腹侧区域的 Aβ 和 tau 的免疫反应性显着增加。可以独立地在切片中诱导 Aβ 斑块和 tau NFT 样病理学。重金属（铝，铅，镉）有力地增强了 Aβ 斑块样病理学，该病理学在细胞死亡之前在细胞内发展。细胞内途径调节剂（东莨菪碱，wortmannin，MHY1485）显着促进了 tau NFT 样病理学。培养基中纳米摩尔浓度的东莨菪碱，wortmannin，MHY1485，铅和镉的组合与未补充培养基的切片相比，强烈增加了腹侧区域的 Aβ 斑块和 tau NFT 样免疫反应性。结果表明，我们可以利用胶原水凝胶传播人 Aβ42 和 P301S 聚集 tau 的潜力，以及药理学操作，产生与 AD 相关的病理学。该结果提供了一种新的离体器官型切片模型，用于研究 AD 病理学，具有未来筛选药物或治疗方法的潜在应用。
