All-Optical Electrophysiology for High-Throughput Functional Characterization of a Human iPSC-Derived Motor Neuron Model of ALS.
All-Optical Electrophysiology for High-Throughput Functional Characterization of a Human iPSC-Derived Motor Neuron Model of ALS.
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DOI：
10.1016/j.stemcr.2018.04.020
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发表时间：
2018-06-05
影响因子：
5.9
通讯作者：
Cohen AE
中科院分区：
文献类型：
--
作者：
Kiskinis E;Kralj JM;Zou P;Weinstein EN;Zhang H;Tsioras K;Wiskow O;Ortega JA;Eggan K;Cohen AE
人类诱导多能干细胞（iPSC）衍生的神经元是一种有吸引力的疾病建模基质，但这些培养物的异质性对通过手动膜片钳电生理学进行功能表征提出了挑战。在这里，我们描述了一个优化的全光电生理学，“Optopatch”管道，用于人类ipsc衍生的神经元培养物的高通量功能表征。我们在肌萎缩性侧索硬化症（ALS）的人类ipsc衍生的运动神经元（iPSC-MN）模型中验证了该方法。在将具有als引起突变（SOD1 A4V）的iPSC-MNs与基因组校正对照进行比较时，突变体在弱或无刺激下显示出更高的峰值速率，并且在强光遗传刺激下更有可能进入去极化区。我们将这些结果与基于简单电导的神经元模型的数值模拟以及基于ipsc的ALS模型的文献结果进行了比较。我们的数据和模拟表明，缓慢激活钾通道的缺陷可能是SOD1 A4V突变中电生理变化的基础。在这篇文章中，Kiskinis和他的同事使用全光学电生理学来表征基于ipsc的ALS模型。通过对来自SOD1 （A4V）突变患者和基因组校正对照患者的运动神经元的兴奋性进行高通量光学测量，作者确定了与突变体中KV7电流缺陷一致的放电差异。
Human induced pluripotent stem cell (iPSC)-derived neurons are an attractive substrate for modeling disease, yet the heterogeneity of these cultures presents a challenge for functional characterization by manual patch-clamp electrophysiology. Here, we describe an optimized all-optical electrophysiology, “Optopatch,” pipeline for high-throughput functional characterization of human iPSC-derived neuronal cultures. We demonstrate the method in a human iPSC-derived motor neuron (iPSC-MN) model of amyotrophic lateral sclerosis (ALS). In a comparison of iPSC-MNs with an ALS-causing mutation (SOD1 A4V) with their genome-corrected controls, the mutants showed elevated spike rates under weak or no stimulus and greater likelihood of entering depolarization block under strong optogenetic stimulus. We compared these results with numerical simulations of simple conductance-based neuronal models and with literature results in this and other iPSC-based models of ALS. Our data and simulations suggest that deficits in slowly activating potassium channels may underlie the changes in electrophysiology in the SOD1 A4V mutation. All-optical electrophysiology enables high-throughput assays in hiPSC-derived neurons Neurons derived from ALS patients fire differently from genome-corrected controls A deficit in the Kv7 potassium current can explain the difference in firing In this article, Kiskinis and coworkers use all-optical electrophysiology to characterize an iPSC-based model of ALS. By performing high-throughput optical measurements of excitability in motor neurons derived from patients with a SOD1 (A4V) mutations and genome-corrected controls, the authors identify differences in firing consistent with a deficit in KV7 current in the mutants.
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影响因子：
8.8
作者：
Wainger BJ;Kiskinis E;Mellin C;Wiskow O;Han SS;Sandoe J;Perez NP;Williams LA;Lee S;Boulting G;Berry JD;Brown RH Jr;Cudkowicz ME;Bean BP;Eggan K;Woolf CJ
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