Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in   single cells

doi:10.1152/ajpcell.00335.2012

CORC > 北京大学 > 生命科学学院

	Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in single cells
	Zheng, Liang-Hong ; Wang, Chang-He ; Shang, Shu-Jiang ; Zhang, Xiao-Yu ; Wang, Ye-Shi ; Wu, Qi-Hui ; Hu, Mei-Qin ; Chai, Zu-Yin ; Wu, Xi ; Zheng, Hui ; Zhang, Chen ; Wang, Lie-Cheng ; Xiong, Wei ; Zhou, Zhuan
刊名	american journal of physiology cell physiology
	2013
关键词	GPCR endocytosis FM imaging DRG neuron confocal microscopy PROTEIN-COUPLED RECEPTOR DORSAL-ROOT GANGLION PERIPHERAL AXOTOMY NEURONS DESENSITIZATION TRAFFICKING INTERNALIZATION ACTIVATION EXPRESSION MEMBRANE
DOI	10.1152/ajpcell.00335.2012
英文摘要	Most G protein-coupled receptors (GPCRs) do not generate membrane currents in response to ligand-receptor binding (LRB). Here, we describe a novel technique using endocytosis as a bioassay that can detect activation of a GPCR in a way analogous to patch-clamp recording of an ion channel in a living cell. The confocal imaging technique, termed FM endocytosis imaging (FEI), can record ligand-GPCR binding with high temporal (second) and spatial (micrometer) resolution. LRB leads to internalization of an endocytic vesicle, which can be labeled by a styryl FM dye and visualized as a fluorescent spot. Distinct from the green fluorescence protein-labeling method, FEI can detect LRB endocytosis mediated by essentially any receptors (GPCRs or receptors of tyrosine kinase) in a native cell/cell line. Three modified versions of FEI permit promising applications in functional GPCR studies and drug screening in living cells: 1) LRB can be recorded in "real time" (time scale of seconds); 2) internalized vesicles mediated by different GPCRs can be discriminated by different colors; and 3) a high throughput method can screen ligands of a specific GPCR.; Cell Biology; Physiology; SCI(E); 0; ARTICLE; 7; C751-C760; 305
语种	英语
内容类型	期刊论文
源URL	[http://ir.pku.edu.cn/handle/20.500.11897/220530]
专题	生命科学学院
推荐引用方式 GB/T 7714	Zheng, Liang-Hong,Wang, Chang-He,Shang, Shu-Jiang,et al. Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in single cells[J]. american journal of physiology cell physiology,2013.
APA	Zheng, Liang-Hong.,Wang, Chang-He.,Shang, Shu-Jiang.,Zhang, Xiao-Yu.,Wang, Ye-Shi.,...&Zhou, Zhuan.(2013).Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in single cells.american journal of physiology cell physiology.
MLA	Zheng, Liang-Hong,et al."Real-time endocytosis imaging as a rapid assay of ligand-GPCR binding in single cells".american journal of physiology cell physiology (2013).