Reconstitution of the Human Nigro-striatal Pathway on-a-Chip Reveals OPA1-Dependent Mitochondrial Defects and Loss of Dopaminergic Synapses
Reconstitution of the Human Nigro-striatal Pathway on-a-Chip Reveals OPA1-Dependent Mitochondrial Defects and Loss of Dopaminergic Synapses
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DOI：
10.1016/j.celrep.2019.11.111
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发表时间：
2019-12-24
期刊：
影响因子：
8.8
通讯作者：
Broccoli, Vania
中科院分区：
文献类型：
--
作者：
Iannielli, Angelo;Ugolini, Giovanni Stefano;Broccoli, Vania
干细胞衍生的神经元通常在缺乏空间组织和任何有意义的连接的大量培养中获得。我们实现了一个微流控系统，用于长期培养具有图案化投射和突触末端的人类神经元。在微芯片上共培养人中脑多巴胺能神经元和纹状体中等多刺神经元，建立了具有功能性多巴胺能突触的协调的黑质-纹状体回路。我们使用这个平台来剖析与具有OPA 1突变的遗传形式的帕金森病（PD）相关的线粒体功能障碍。值得注意的是，我们发现OPA 1突变多巴胺能神经元的轴突表现出线粒体质量的显着减少。这种缺陷会导致多巴胺能突触的显著丧失，这在长期培养中是不可能的。因此，PD相关的突触线粒体耗竭可能先于神经元连接丧失和神经变性。通过微流控技术在体外重建人类回路，通过建立有序的神经元区室和正确的突触身份，为研究大脑网络提供了一个强大的系统。
Stem cell-derived neurons are generally obtained in mass cultures that lack both spatial organization and any meaningful connectivity. We implement a microfluidic system for long-term culture of human neurons with patterned projections and synaptic terminals. Co-culture of human midbrain dopaminergic and striatal medium spiny neurons on the microchip establishes an orchestrated nigro-striatal circuitry with functional dopaminergic synapses. We use this platform to dissect the mitochondrial dysfunctions associated with a genetic form of Parkinson's disease (PD) with OPA1 mutations. Remarkably, we find that axons of OPA1 mutant dopaminergic neurons exhibit a significant reduction of mitochondrial mass. This defect causes a significant loss of dopaminergic synapses, which worsens in long-term cultures. Therefore, PD-associated depletion of mitochondria at synapses might precede loss of neuronal connectivity and neurodegeneration. In vitro reconstitution of human circuitries by microfluidic technology offers a powerful system to study brain networks by establishing ordered neuronal compartments and correct synapse identity.