Reactive Oxygen Species Mediate Transcriptional Responses to Dopamine and Cocaine in Human Cerebral Organoids.
Reactive Oxygen Species Mediate Transcriptional Responses to Dopamine and Cocaine in Human Cerebral Organoids.
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活性氧介导人类大脑类器官对多巴胺和可卡因的转录反应。
DOI：
10.1101/2023.06.13.544782
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发表时间：
2023
期刊：
影响因子：
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通讯作者：
Keung,AlbertJ
中科院分区：
文献类型：
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作者：
Rudibaugh,ThomasT;Keung,AlbertJ
多巴胺信号在成人腹侧前脑调节行为、应激反应和记忆形成，在神经发育中调节神经分化和细胞迁移。过量的多巴胺水平，包括在子宫内和成人中因使用可卡因而导致的多巴胺水平，可能导致长期的不良后果。体内平衡和病理变化的机制尚不清楚，部分原因是多巴胺引起的细胞反应不同，并且依赖于多巴胺信号的物种特异性差异的动物模型。在这项研究中，我们使用人类来源的腹侧前脑类器官模型，并表征它们对可卡因或多巴胺的反应。我们探索多巴胺或可卡因的剂量方案来模拟急性或慢性暴露。然后，我们使用钙成像、cAMP成像和大量rna测序来测量对可卡因或多巴胺暴露的反应。我们观察到除了氧化应激指标暴露后炎症途径的上调。利用活性氧（ROS）抑制剂，我们证明了ROS对于可卡因暴露的多重转录反应是必要的。这些结果强调了新的反应途径，并验证了脑类器官作为研究大脑复杂生物过程的体外人体模型的潜力。
Dopamine signaling in the adult ventral forebrain regulates behavior, stress response, and memory formation and in neurodevelopment regulates neural differentiation and cell migration. Excessive dopamine levels, including those due to cocaine use in utero and in adults, could lead to long-term adverse consequences. The mechanisms underlying both homeostatic and pathological changes remain unclear, in part due to the diverse cellular responses elicited by dopamine and the reliance on animal models that exhibit species-specific differences in dopamine signaling. In this study, we use the human-derived ventral forebrain organoid model of Xiang–Tanaka and characterize their response to cocaine or dopamine. We explore dosing regimens of dopamine or cocaine to simulate acute or chronic exposure. We then use calcium imaging, cAMP imaging, and bulk RNA-sequencing to measure responses to cocaine or dopamine exposure. We observe an upregulation of inflammatory pathways in addition to indicators of oxidative stress following exposure. Using inhibitors of reactive oxygen species (ROS), we then show ROS to be necessary for multiple transcriptional responses of cocaine exposure. These results highlight novel response pathways and validate the potential of cerebral organoids as in vitro human models for studying complex biological processes in the brain.