Related Experiment Video
Updated: Jun 3, 2026
16:16
Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
Published on: September 13, 2013
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A molecular toolbox to study progesterone receptor signaling
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
Summary
New tools enable sensitive measurement of progesterone receptor (PR) signaling in breast cells. These reporters quantify PR activity and visualize pathway dynamics, aiding breast cancer research.
Area of Science:
- Cell biology
- Endocrinology
- Cancer research
Background:
- Progesterone receptor (PR) signaling is crucial for mammary gland development and homeostasis.
- Studying PR signaling is challenging due to a lack of sensitive quantitative and spatial assays.
- Human breast epithelial cells are key models for understanding PR-mediated processes.
Purpose of the Study:
- To develop novel tools for sensitive measurement and visualization of PR signaling in human breast epithelial cells.
- To establish reporter assays for quantifying PR pathway activity across various progesterone concentrations.
- To enable single-cell level visualization of PR signaling dynamics.
Main Methods:
- Generation of optimized Progesterone Responsive Element (PRE)-luciferase and PRE-green fluorescent protein (GFP) lentiviral reporter constructs.
- Quantification of PR signaling activity in MCF7 and T47D breast cancer cell lines using luciferase assays.
- Visualization of PR signaling at the single-cell level using fluorescent reporters.
- Assessment of PR expression levels and their impact on reporter assay sensitivity and endogenous gene induction via qRT-PCR.
Main Results:
- Optimized PRE-luciferase reporters accurately quantify PR signaling across a wide range of progesterone concentrations.
- A fluorescent lentiviral PRE-GFP reporter enables visualization of PR signaling at the single-cell level.
- Increased PR expression significantly enhances the dynamic range of reporter assays and endogenous PR target gene induction.
- PR signaling response exhibits variability based on cell line, target gene, and hormone concentration.
Conclusions:
- Developed novel reporter tools provide sensitive and versatile methods for measuring PR signaling in breast epithelial cells.
- Enhanced PR expression is critical for robust PR signaling assays.
- These tools facilitate a more rationally designed approach to studying PR signaling in breast biology and disease.
Related Concept Videos
01:21G-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.
01:08Intracellular 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
01:21G-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.
01:15G 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...
01:15G 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...
01:26Secondary 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...