Alich Therese C, Röderer Pascal, Szalontai Balint, Golcuk Kurt, Tariq Shahan, Peitz Michael, Brüstle Oliver, Mody Istvan
Institute of Experimental Epileptology and Cognition Research, Medical Faculty, University Hospital Bonn, Bonn, Germany.
Institute of Reconstructive Neurobiology, Medical Faculty, University Hospital Bonn, Bonn, Germany.
Front Cell Neurosci. 2023 Jan 17;16:1039957. doi: 10.3389/fncel.2022.1039957. eCollection 2022.
Human induced pluripotent stem cells (hiPSCs) are a promising approach to study neurological and neuropsychiatric diseases. Most methods to record the activity of these cells have major drawbacks as they are invasive or they do not allow single cell resolution. Genetically encoded voltage indicators (GEVIs) open the path to high throughput visualization of undisturbed neuronal activity. However, conventional GEVIs perturb membrane integrity through inserting multiple copies of transmembrane domains into the plasma membrane. To circumvent large add-ons to the plasma membrane, we used a minimally invasive novel hybrid dark quencher GEVI to record the physiological and pathological firing patterns of hiPSCs-derived sensory neurons from patients with inherited erythromelalgia, a chronic pain condition associated with recurrent attacks of redness and swelling in the distal extremities. We observed considerable differences in action potential firing patterns between patient and control neurons that were previously overlooked with other recording methods. Our system also performed well in hiPSC-derived forebrain neurons where it detected spontaneous synchronous bursting behavior, thus opening the path to future applications in other cell types and disease models including Parkinson's disease, Alzheimer's disease, epilepsy, and schizophrenia, conditions associated with disturbances of neuronal activity and synchrony.
人类诱导多能干细胞(hiPSC)是研究神经和神经精神疾病的一种很有前景的方法。大多数记录这些细胞活性的方法都有主要缺点,因为它们具有侵入性,或者无法实现单细胞分辨率。基因编码电压指示剂(GEVI)为未受干扰的神经元活动的高通量可视化开辟了道路。然而,传统的GEVI通过将多个跨膜结构域拷贝插入质膜来扰乱膜的完整性。为了避免对质膜进行大量附加,我们使用了一种微创新型混合暗猝灭剂GEVI来记录遗传性红斑性肢痛症患者的hiPSC衍生感觉神经元的生理和病理放电模式,遗传性红斑性肢痛症是一种慢性疼痛疾病,与远端肢体反复出现的红肿发作有关。我们观察到患者神经元和对照神经元之间的动作电位放电模式存在显著差异,而这些差异在以前的其他记录方法中被忽视了。我们的系统在hiPSC衍生的前脑神经元中也表现良好,在那里它检测到了自发同步爆发行为,从而为未来在其他细胞类型和疾病模型(包括帕金森病、阿尔茨海默病、癫痫和精神分裂症,这些疾病都与神经元活动和同步性紊乱有关)中的应用开辟了道路。