Notch信号通路
Notch蛋白质类
细胞生物学
Hes3信号轴
生物
转录因子
信号转导
受体
活体细胞成像
细胞命运测定
细胞
基因
遗传学
作者
Emil M. Hansson,Ana I. Teixeira,Maria Gustafsson,Takeaki Dohda,Gavin Chapman,Konstantinos Meletis,Jonas Muhr,Urban Lendahl
出处
期刊:Developmental Neuroscience
[S. Karger AG]
日期:2006-01-01
卷期号:28 (1-2): 118-127
被引量:43
摘要
Notch signaling is a highly conserved signaling pathway, which is critical for many cell fate decisions. Ligand activation of Notch leads to cleavage of the Notch receptor and liberation of the Notch intracellular domain (ICD) from the membrane-tethered receptor. After translocation to the nucleus, the Notch ICD interacts with the DNA-binding protein CSL to activate gene transcription. To better understand the temporal and spatial aspects of Notch signaling, we here describe a fluorescent protein-based reporter assay that allows Notch activation to be followed in real time in individual cells. We have generated a reporter construct composed of 12 CSL-binding motifs linked to fluorescent proteins with different half-lives: a stabler red fluorescent protein (DsRedExpressDR) and a destabilized form of green fluorescent protein (d1EGFP). The fluorescent reporters reflect the activation status of Notch signaling with single-cell resolution. The reporters rapidly respond to various forms of Notch activation, including ligand activation of full-length Notch receptors. Finally, we use this assay to gain insights into the level of Notch signaling in CNS progenitor cells in culture and in vivo.
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