Reactive Astrocytes Contribute to Alzheimer's Disease-Related Neurotoxicity and Synaptotoxicity in a Neuron-Astrocyte Co-culture Assay.
Reactive Astrocytes Contribute to Alzheimer's Disease-Related Neurotoxicity and Synaptotoxicity in a Neuron-Astrocyte Co-culture Assay.
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DOI：
10.3389/fncel.2021.739411
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发表时间：
2021
影响因子：
5.3
通讯作者：
Krasemann S
中科院分区：
文献类型：
--
作者：
Wasilewski D;Villalba-Moreno ND;Stange I;Glatzel M;Sepulveda-Falla D;Krasemann S
阿尔茨海默病（AD）的病理特征包括淀粉样蛋白- β (Aβ)的沉积和积累，神经原纤维缠结的形成和神经元的丢失。症状前疾病阶段的发病机制仍然难以捉摸，尽管研究表明早期结构和功能改变可能发生在神经元树突棘。症状前的改变也可能影响不同的中枢神经系统细胞类型。然而，这些细胞类型作为病理原因或结果的具体贡献很难在体内研究。目前缺乏相对简单、定义明确且经过验证的体外模型，这些模型可以直接解释结果并概括病理生理学的各个方面。例如，解剖ad相关过程（例如，神经毒性与突触毒性）可能很难与普通细胞为基础的系统，如神经细胞系或原代神经元。为了研究和表征反应性星形胶质细胞在ad相关提示下对神经元形态的影响，我们在所谓的Banker“三明治”共培养实验的基础上修改了小鼠原代神经元和小鼠原代星形胶质细胞的体外共培养实验。在这里，我们提供了一个简单和模块化的实验，用完全分化的小鼠原代神经元来研究神经元和星形胶质细胞在共培养环境下的旁分泌相互作用。在我们的实验中，两种细胞类型的读数都得到了。星形胶质细胞饲养细胞通过Aβ42、Aβ40或脂多糖（LPS）预先暴露于神经炎症条件下。反应性星形胶质细胞对神经元的非细胞自主毒性作用使用Sholl分析来评估树突复杂性，而突触点作为突触毒性的读数。在这里，我们发现星形胶质细胞在AD特异性背景下积极地促进了原代神经元的表型，强调了不同细胞类型在AD病理中的作用。星形胶质细胞中细胞因子的表达模式明显改变。值得注意的是，反应性星形胶质细胞对神经元的影响高度依赖于定义的细胞比例。我们的共培养系统是模块化的，成本低，并允许我们在明确的实验条件下探测两种主要中枢神经系统细胞类型（神经元和星形胶质细胞）之间的神经变性和神经炎症方面。我们易于遵循的方案，包括工作流程图，也可能为未来在其他疾病背景下研究星形胶质细胞和神经元的相互作用提供方法学大纲。
Pathological hallmarks of Alzheimer’s disease (AD) include deposition and accumulation of amyloid- β (Aβ), neurofibrillary tangle formation, and neuronal loss. Pathogenesis of presymptomatic disease stages remains elusive, although studies suggest that the early structural and functional alterations likely occur at neuronal dendritic spines. Presymptomatic alterations may also affect different CNS cell types. However, specific contributions of these cell types as cause or consequence of pathology are difficult to study in vivo. There is a shortage of relatively simple, well-defined, and validated in vitro models that allow a straightforward interpretation of results and recapitulate aspects of pathophysiology. For instance, dissecting the AD-related processes (e.g., neurotoxicity vs. synaptotoxicity) may be difficult with the common cell-based systems such as neuronal cell lines or primary neurons. To investigate and characterize the impact of reactive astrocytes on neuronal morphology in the context of AD-related cues, we modified an in vitro co-culture assay of primary mouse neurons and primary mouse astrocytes based on the so-called Banker “sandwich” co-culture assay. Here, we provide a simple and modular assay with fully differentiated primary mouse neurons to study the paracrine interactions between the neurons and the astrocytes in the co-culture setting. Readouts were obtained from both cell types in our assay. Astrocyte feeder cells were pre-exposed to neuroinflammatory conditions by means of Aβ42, Aβ40, or lipopolysaccharide (LPS). Non-cell autonomous toxic effects of reactive astrocytes on neurons were assessed using the Sholl analysis to evaluate the dendritic complexity, whereas synaptic puncta served as a readout of synaptotoxicity. Here, we show that astrocytes actively contribute to the phenotype of the primary neurons in an AD-specific context, emphasizing the role of different cell types in AD pathology. The cytokine expression pattern was significantly altered in the treated astrocytes. Of note, the impact of reactive astrocytes on neurons was highly dependent on the defined cell ratios. Our co-culture system is modular, of low cost, and allows us to probe aspects of neurodegeneration and neuroinflammation between the two major CNS cell types, neurons, and astrocytes, under well-defined experimental conditions. Our easy-to-follow protocol, including work-flow figures, may also provide a methodological outline to study the interactions of astrocytes and neurons in the context of other diseases in the future.
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DOI：
10.1016/j.trci.2018.04.008
发表时间：
2018
期刊：
Alzheimer's & dementia (New York, N. Y.)
影响因子：
--
作者：
Cummings J;Reiber C;Kumar P
通讯作者：
Kumar P
影响因子：
14.8
作者：
Biederer, Thomas;Scheiffele, Peter
通讯作者：
Scheiffele, Peter
影响因子：
14.8
作者：
Beaudoin, Gerard M. J., III;Lee, Seung-Hye;Arikkath, Jyothi
通讯作者：
Arikkath, Jyothi
DOI：
10.1007/978-1-61779-452-0_22
发表时间：
2012-01-01
期刊：
ASTROCYTES: METHODS AND PROTOCOLS
影响因子：
--
作者：
Jones, Emma V.;Cook, Denise;Murai, Keith K.
通讯作者：
Murai, Keith K.
影响因子：
4.7
作者：
Falker, Clemens;Hartmann, Alexander;Glatzel, Markus
通讯作者：
Glatzel, Markus