Article ; Online: Characterization of a far-red analog of ghrelin for imaging GHS-R in P19-derived cardiomyocytes.
2014 Volume 54, Page(s) 81–88
Abstract: Ghrelin and its receptor, the growth hormone secretagogue receptor (GHS-R), are expressed ... of determining GHS-R expression in situ. We now report the generation and characterization of a far-red ghrelin ... in GHS-R in developing cardiomyocytes. We also generated the des-acyl analog, des-acyl [Lys(19)(Cy5 ...
| Abstract | Ghrelin and its receptor, the growth hormone secretagogue receptor (GHS-R), are expressed in the heart, and may function to promote cardiomyocyte survival, differentiation and contractility. Previously, we had generated a truncated analog of ghrelin conjugated to fluorescein isothiocyanate for the purposes of determining GHS-R expression in situ. We now report the generation and characterization of a far-red ghrelin analog, [Dpr(3)(octanoyl), Lys(19)(Cy5)]ghrelin (1-19), and show that it can be used to image changes in GHS-R in developing cardiomyocytes. We also generated the des-acyl analog, des-acyl [Lys(19)(Cy5)]ghrelin (1-19) and characterized its binding to mouse heart sections. Receptor binding affinity of Cy5-ghrelin as measured in HEK293 cells overexpressing GHS-R1a was within an order of magnitude of that of fluorescein-ghrelin and native human ghrelin, while the des-acyl Cy5-ghrelin did not bind GHS-R1a. Live cell imaging in HEK293/GHS-R1a cells showed cell surface labeling that was displaced by excess ghrelin. Interestingly, Cy5-ghrelin, but not the des-acyl analog, showed concentration-dependent binding in mouse heart tissue sections. We then used Cy5-ghrelin to track GHS-R expression in P19-derived cardiomyocytes. Live cell imaging at different time points after DMSO-induced differentiation showed that GHS-R expression preceded that of the differentiation marker aMHC and tracked with the contractility marker SERCA 2a. Our far-red analog of ghrelin adds to the tools we are developing to map GHS-R in developing and diseased cardiac tissues. |
|---|---|
| MeSH term(s) | Amino Acid Sequence ; Animals ; Cell Differentiation ; Female ; Ghrelin/analogs & derivatives ; Ghrelin/metabolism ; HEK293 Cells/metabolism ; Humans ; Male ; Mice, Inbred C57BL ; Molecular Imaging/methods ; Molecular Sequence Data ; Molecular Structure ; Myocardium/cytology ; Myocardium/metabolism ; Myocytes, Cardiac/cytology ; Myocytes, Cardiac/metabolism ; Peptide Fragments ; Receptors, Ghrelin/analysis ; Receptors, Ghrelin/metabolism |
| Chemical Substances | (Dpr3(octanoyl)Lys19(Cy5))ghrelin(1-19) ; Ghrelin ; Peptide Fragments ; Receptors, Ghrelin ; ghrelin, des-n-octanoyl |
| Language | English |
| Publishing date | 2014-04 |
| Publishing country | United States |
| Document type | Journal Article ; Research Support, Non-U.S. Gov't |
| ZDB-ID | 769028-9 |
| ISSN | 1873-5169 ; 0196-9781 |
| ISSN (online) | 1873-5169 |
| ISSN | 0196-9781 |
| DOI | 10.1016/j.peptides.2014.01.011 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
More links
Kategorien
In stock of ZB MED Cologne/Königswinter
| Zs.A 1649: Show issues | Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG) |
Order via subito
This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.