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通过时间身份因素直接进行神经元重编程。

Direct neuronal reprogramming by temporal identity factors.

机构信息

Cellular Neurobiology Research Unit, Montreal Clinical Research Institute (IRCM), Montreal, QC H2W 1R7, Canada.

Integrated Program in Neuroscience, McGill University, Montreal, QC H3A 1A1, Canada.

出版信息

Proc Natl Acad Sci U S A. 2023 May 9;120(19):e2122168120. doi: 10.1073/pnas.2122168120. Epub 2023 May 1.

Abstract

Temporal identity factors are sufficient to reprogram developmental competence of neural progenitors and shift cell fate output, but whether they can also reprogram the identity of terminally differentiated cells is unknown. To address this question, we designed a conditional gene expression system that allows rapid screening of potential reprogramming factors in mouse retinal glial cells combined with genetic lineage tracing. Using this assay, we found that coexpression of the early temporal identity transcription factors Ikzf1 and Ikzf4 is sufficient to directly convert Müller glial (MG) cells into cells that translocate to the outer nuclear layer (ONL), where photoreceptor cells normally reside. We name these "induced ONL (iONL)" cells. Using genetic lineage tracing, histological, immunohistochemical, and single-cell transcriptome and multiome analyses, we show that expression of Ikzf1/4 in MG in vivo, without retinal injury, mostly generates iONL cells that share molecular characteristics with bipolar cells, although a fraction of them stain for Rxrg, a cone photoreceptor marker. Furthermore, we show that coexpression of Ikzf1 and Ikzf4 can reprogram mouse embryonic fibroblasts to induced neurons in culture by rapidly remodeling chromatin and activating a neuronal gene expression program. This work uncovers general neuronal reprogramming properties for temporal identity factors in terminally differentiated cells.

摘要

时间身份因素足以重新编程神经祖细胞的发育能力并改变细胞命运输出,但它们是否也能重新编程终末分化细胞的身份尚不清楚。为了解决这个问题,我们设计了一种条件基因表达系统,允许在小鼠视网膜神经胶质细胞中快速筛选潜在的重编程因子,并结合遗传谱系追踪。使用该测定法,我们发现早期时间身份转录因子 Ikzf1 和 Ikzf4 的共表达足以直接将 Muller 胶质细胞 (MG) 细胞转化为迁移到外核层 (ONL) 的细胞,感光细胞通常位于该层。我们将这些细胞命名为“诱导的 ONL (iONL)”细胞。通过遗传谱系追踪、组织学、免疫组织化学以及单细胞转录组和多组学分析,我们表明,在体内无视网膜损伤的情况下,MG 中 Ikzf1/4 的表达主要产生与双极细胞具有分子特征的 iONL 细胞,尽管其中一部分会对 Rxrg 染色,Rxrg 是一种圆锥状光感受器标记物。此外,我们表明 Ikzf1 和 Ikzf4 的共表达可以通过快速重塑染色质并激活神经元基因表达程序,将小鼠胚胎成纤维细胞在培养中重编程为诱导神经元。这项工作揭示了时间身份因素在终末分化细胞中进行普遍神经元重编程的特性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ab92/10175841/1baefd1afc8d/pnas.2122168120fig01.jpg

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