Proper brain function relies on the precise arrangement and flow of information between diverse neural subtypes. Developing improved human cell-based models which faithfully mimic biologically relevant connectivity patterns may improve drug screening efforts given the limited success of animal models to predict safety and efficacy of therapeutics in human clinical trials. To address this need, we have developed experimental models of defined neural circuitries through the compartmentalization of neuronal cell subtypes in a 96 well plate-based platform where each microwell is divided into two compartments connected by microchannels allowing high-throughput screening (HTS) of small molecules. We demonstrate that we can generate subtype-specific excitatory and inhibitory induced neuronal cells (iNs) from human stem cell lines and that these neurons form robust functional circuits with defined connectivity. Through the use of the genetically encoded calcium indicator GCaMP6f, we monitor calcium ion transients generated during neuronal firing between and within compartments. We further demonstrate functionality of the circuit by perturbing network activity through the addition of glutamate receptor blockers using automated liquid handling. Lastly, we show that we can stimulate network activity in defined neuronal subtypes through the expression of the designer receptor exclusively activated by designer drugs (DREADD) hM3Dq and application of the ligand clozapine-N-oxide (CNO). Our results demonstrate the formation of functional neural circuits in a high-throughput platform that is compatible with compound screening, representing an important step towards developing new screening platforms for studying and ultimately treating psychiatric brain disorders that arise from disordered neural circuit function.

中文翻译：

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使用人类诱导神经元开发用于药物筛选应用的高通量阵列神经电路平台。

正确的大脑功能依赖于不同神经亚型之间信息的精确排列和流动。鉴于动物模型在人类临床试验中预测治疗方法的安全性和有效性的成功有限，开发改进的基于人类细胞的模型，忠实地模拟生物相关的连接模式，可能会改善药物筛选工作。为了满足这一需求，我们通过在基于 96 孔板的平台中划分神经元细胞亚型，开发了定义神经回路的实验模型，其中每个微孔分为两个由微通道连接的隔室，允许高通量筛选 (HTS)分子。我们证明我们可以从人类干细胞系生成亚型特异性的兴奋性和抑制性诱导神经元细胞 (iNs)，并且这些神经元形成具有明确连接性的强大功能回路。通过使用基因编码的钙指示剂 GCaMP6f，我们监测在隔间之间和隔间内的神经元放电过程中产生的钙离子瞬变。我们通过使用自动液体处理添加谷氨酸受体阻滞剂来扰乱网络活动，进一步证明了电路的功能。最后，我们表明我们可以通过表达由设计药物 (DREADD) hM3Dq 专门激活的设计受体和应用配体氯氮平-N-氧化物 (CNO) 来刺激特定神经元亚型中的网络活动。