Back to Search
Start Over
The use of fluorescence correlation spectroscopy to monitor cell surface β2‐adrenoceptors at low expression levels in human embryonic stem cell‐derived cardiomyocytes and fibroblasts
- Source :
- The FASEB Journal. 35
- Publication Year :
- 2021
- Publisher :
- Wiley, 2021.
-
Abstract
- The importance of cell phenotype in determining the molecular mechanisms underlying β2 -adrenoceptor (β2AR) function has been noted previously when comparing responses in primary cells and recombinant model cell lines. Here, we have generated haplotype-specific SNAP-tagged β2AR human embryonic stem (ES) cell lines and applied fluorescence correlation spectroscopy (FCS) to study cell surface receptors in progenitor cells and in differentiated fibroblasts and cardiomyocytes. FCS was able to quantify SNAP-tagged β2AR number and diffusion in both ES-derived cardiomyocytes and CRISPR/Cas9 genome-edited HEK293T cells, where the expression level was too low to detect using standard confocal microscopy. These studies demonstrate the power of FCS in investigating cell surface β2ARs at the very low expression levels often seen in endogenously expressing cells. Furthermore, the use of ES cell technology in combination with FCS allowed us to demonstrate that cell surface β2ARs internalize in response to formoterol-stimulation in ES progenitor cells but not following their differentiation into ES-derived fibroblasts. This indicates that the process of agonist-induced receptor internalization is strongly influenced by cell phenotype and this may have important implications for drug treatment with long-acting β2AR agonists.
- Subjects :
- 0301 basic medicine
Cell
Biochemistry
law.invention
03 medical and health sciences
0302 clinical medicine
Cell surface receptor
Confocal microscopy
law
Genetics
medicine
Humans
Myocytes, Cardiac
Progenitor cell
Primary cell
Adrenergic beta-2 Receptor Agonists
Molecular Biology
Embryonic Stem Cells
Fluorescent Dyes
Chemistry
HEK 293 cells
Membrane Proteins
Cell Differentiation
Fibroblasts
Propranolol
Embryonic stem cell
Cell biology
HEK293 Cells
Spectrometry, Fluorescence
030104 developmental biology
medicine.anatomical_structure
Gene Expression Regulation
Cell culture
Receptors, Adrenergic, beta-2
030217 neurology & neurosurgery
Biotechnology
Subjects
Details
- ISSN :
- 15306860 and 08926638
- Volume :
- 35
- Database :
- OpenAIRE
- Journal :
- The FASEB Journal
- Accession number :
- edsair.doi.dedup.....28b4acb5a88296c4c5ff56dd129add3b