一个分子工具箱来研究孕激素受体信号传递
Marleen T Aarts1, Muriel Wagner1, Tanne van der Wal1
1Developmental, Stem Cell and Cancer Biology, Swammerdam Institute for Life Sciences, University of Amsterdam, Science Park 904, 1098 XH, Amsterdam, the Netherlands.
Journal of mammary gland biology and neoplasia
|November 29, 2023
概括
新的工具可以对乳腺细胞中孕激素受体 (PR) 信号的敏感测量. 这些记者量化公关活动和可视化路径动态,帮助乳腺癌研究.
科学领域:
- 细胞生物学 细胞生物学
- 内分泌学 在内分泌学.
- 癌症研究 癌症研究
背景情况:
- 孕激素受体 (PR) 信号传递对于乳腺发育和恒温至关重要.
- 由于缺乏敏感的定量和空间分析,研究PR信号具有挑战性.
- 人类乳腺上皮细胞是理解PR介导过程的关键模型.
研究的目的:
- 开发用于敏感测量和可视化人类乳腺上皮细胞中PR信号的新工具.
- 建立报告测试以量化PR通路活性在不同度的孕激素.
- 为了使PR信号动态的单个细胞水平可视化.
主要方法:
- 优化的孕激素反应元件 (PRE) - 化酶和PRE-绿色光蛋白 (GFP) 乳腺病毒记者结构的生成.
- 使用光酶试验量化MCF7和T47D乳腺癌细胞系中的PR信号活动.
- 使用光报告器在单细胞水平上可视化PR信号.
- 评估PR表达水平及其对记者测定灵敏度及其通过qRT-PCR对内源基因诱导的影响.
主要成果:
- 优化的PRE-luciferase记者可以准确量化PR信号,跨越广泛的孕激素度.
- 一个光透镜病毒 PRE-GFP 报告员使得在单细胞水平上可视化 PR 信号.
- 增加PR表达显著提高了记者测定和内源性PR基因诱导的动态范围.
- PR信号响应表现出基于细胞系,基因和激素度的变异性.
结论:
- 开发的新型报告工具为测量乳腺上皮细胞中的PR信号提供了灵敏和多功能方法.
- 增强的PR表达对于强大的PR信号测试至关重要.
- 这些工具促进了一种更合理的设计方法来研究乳腺生物学和疾病中的PR信号.
相关概念视频
G-protein Coupled Receptors
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
Intracellular Hormone Receptors
Lipid-soluble hormones diffuse across the plasma and nuclear membrane of target cells to bind to their specific intracellular receptors. These receptors act as transcription factors that regulate gene expression and protein synthesis in the target cell
G-protein Coupled Receptors
G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
G Protein-coupled Receptors
G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
G Protein-coupled Receptors
G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
Secondary Messengers in Hormone Action
Water-soluble hormones cannot cross the plasma membrane, so they rely on protein receptors that span the membrane to trigger intracellular signaling pathways. These pathways then activate second messengers inside the cell, including cAMP or calcium ions.
Many hormones bind to transmembrane G protein-coupled receptors that connect to regulatory G proteins. These G proteins can then activate enzymes such as adenylyl cyclase or phospholipase C. Adenylyl cyclase converts ATP to cAMP, activating...
Many hormones bind to transmembrane G protein-coupled receptors that connect to regulatory G proteins. These G proteins can then activate enzymes such as adenylyl cyclase or phospholipase C. Adenylyl cyclase converts ATP to cAMP, activating...


