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诱导多巴胺能神经元用于特定神经亚型的精神疾病风险建模。

Induction of dopaminergic neurons for neuronal subtype-specific modeling of psychiatric disease risk.

机构信息

Pamela Sklar Division of Psychiatric Genomics, Department of Genetics and Genomics, Icahn Institute of Genomics and Multiscale Biology, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

Nash Family Department of Neuroscience, Friedman Brain Institute, Icahn School of Medicine at Mount Sinai, New York, NY, USA.

出版信息

Mol Psychiatry. 2023 May;28(5):1970-1982. doi: 10.1038/s41380-021-01273-0. Epub 2021 Sep 7.

Abstract

Dopaminergic neurons are critical to movement, mood, addiction, and stress. Current techniques for generating dopaminergic neurons from human induced pluripotent stem cells (hiPSCs) yield heterogenous cell populations with variable purity and inconsistent reproducibility between donors, hiPSC clones, and experiments. Here, we report the rapid (5 weeks) and efficient (~90%) induction of induced dopaminergic neurons (iDANs) through transient overexpression of lineage-promoting transcription factors combined with stringent selection across five donors. We observe maturation-dependent increase in dopamine synthesis and electrophysiological properties consistent with midbrain dopaminergic neuron identity, such as slow-rising after- hyperpolarization potentials, an action potential duration of ~3 ms, tonic sub-threshold oscillatory activity, and spontaneous burst firing at a frequency of ~1.0-1.75 Hz. Transcriptome analysis reveals robust expression of genes involved in fetal midbrain dopaminergic neuron identity. Specifically expressed genes in iDANs, as well as those from isogenic induced GABAergic and glutamatergic neurons, were enriched in loci conferring heritability for cannabis use disorder, schizophrenia, and bipolar disorder; however, each neuronal subtype demonstrated subtype-specific heritability enrichments in biologically relevant pathways, and iDANs alone were uniquely enriched in autism spectrum disorder risk loci. Therefore, iDANs provide a critical tool for modeling midbrain dopaminergic neuron development and dysfunction in psychiatric disease.

摘要

多巴胺能神经元对于运动、情绪、成瘾和应激至关重要。目前,从人诱导多能干细胞(hiPSC)生成多巴胺能神经元的技术产生了具有不同纯度的异质细胞群体,并且在供体、hiPSC 克隆和实验之间存在不一致的可重复性。在这里,我们报告了通过瞬时过表达谱系促进转录因子与严格的选择相结合,快速(5 周)和高效(~90%)诱导诱导多巴胺能神经元(iDAN)。我们观察到多巴胺合成和电生理特性与中脑多巴胺能神经元身份一致的成熟依赖性增加,例如慢上升的后超极化电位、约 3 ms 的动作电位持续时间、紧张阈下振荡活动和自发爆发放电频率约为 1.0-1.75 Hz。转录组分析显示与胎儿中脑多巴胺能神经元身份相关的基因的表达稳健。iDAN 中特异性表达的基因,以及与同源诱导的 GABA 能和谷氨酸能神经元中特异性表达的基因,在大麻使用障碍、精神分裂症和双相情感障碍的遗传易感性相关基因座中富集;然而,每个神经元亚型在生物学上相关的途径中都表现出特定的遗传易感性富集,而 iDAN 仅在自闭症谱系障碍风险基因座中富集。因此,iDAN 为在精神疾病中模拟中脑多巴胺能神经元发育和功能障碍提供了重要工具。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6301/8898985/3af568b44e22/nihms-1734801-f0001.jpg

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