Assetta Benedetta, Tang Changyong, Bian Jing, O'Rourke Ryan, Connolly Kevin, Brickler Thomas, Chetty Sundari, Huang Yu-Wen Alvin
Department of Molecular Biology, Cell Biology and Biochemistry, Brown University.
Department of Molecular Biology, Cell Biology and Biochemistry, Brown University; Department of Neurology, The Third Affiliated Hospital of Sun Yat-Sen University.
J Vis Exp. 2020 Nov 9(165). doi: 10.3791/61778.
In Alzheimer's disease (AD) and other neurodegenerative disorders, oligodendroglial failure is a common early pathological feature, but how it contributes to disease development and progression, particularly in the gray matter of the brain, remains largely unknown. The dysfunction of oligodendrocyte lineage cells is hallmarked by deficiencies in myelination and impaired self-renewal of oligodendrocyte precursor cells (OPCs). These two defects are caused at least in part by the disruption of interactions between neuron and oligodendrocytes along the buildup of pathology. OPCs give rise to myelinating oligodendrocytes during CNS development. In the mature brain cortex, OPCs are the major proliferative cells (comprising ~5% of total brain cells) and control new myelin formation in a neural activity-dependent manner. Such neuron-to-oligodendrocyte communications are significantly understudied, especially in the context of neurodegenerative conditions such as AD, due to the lack of appropriate tools. In recent years, our group and others have made significant progress to improve currently available protocols to generate functional neurons and oligodendrocytes individually from human pluripotent stem cells. In this manuscript, we describe our optimized procedures, including the establishment of a co-culture system to model the neuron-oligodendrocyte connections. Our illustrative results suggest an unexpected contribution from OPCs/oligodendrocytes to the brain amyloidosis and synapse integrity and highlight the utility of this methodology for AD research. This reductionist approach is a powerful tool to dissect the specific hetero-cellular interactions out of the inherent complexity inside the brain. The protocols we describe here are expected to facilitate future studies on oligodendroglial defects in the pathogenesis of neurodegeneration.
在阿尔茨海默病(AD)和其他神经退行性疾病中,少突胶质细胞功能障碍是一种常见的早期病理特征,但它如何促进疾病的发展和进展,尤其是在大脑灰质中,在很大程度上仍不清楚。少突胶质细胞谱系细胞的功能障碍以髓鞘形成缺陷和少突胶质前体细胞(OPC)自我更新受损为特征。这两个缺陷至少部分是由病理形成过程中神经元与少突胶质细胞之间相互作用的破坏引起的。在中枢神经系统发育过程中,OPC可分化为形成髓鞘的少突胶质细胞。在成熟的大脑皮层中,OPC是主要的增殖细胞(约占脑细胞总数的5%),并以神经活动依赖的方式控制新的髓鞘形成。由于缺乏合适的工具,这种神经元与少突胶质细胞之间的通讯在很大程度上未得到充分研究,尤其是在AD等神经退行性疾病的背景下。近年来,我们团队和其他团队在改进现有方案以从人多能干细胞中分别生成功能性神经元和少突胶质细胞方面取得了重大进展。在本手稿中,我们描述了我们优化的程序,包括建立一个共培养系统来模拟神经元与少突胶质细胞的连接。我们的说明性结果表明,OPC/少突胶质细胞对脑淀粉样变性和突触完整性有意外贡献,并突出了该方法在AD研究中的实用性。这种简化方法是一种强大的工具,可从大脑固有的复杂性中剖析特定的异细胞相互作用。我们在此描述的方案有望促进未来对神经退行性疾病发病机制中少突胶质细胞缺陷的研究。