In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor
Abstract The Apelin receptor (APLNR), a class A G-protein coupled receptor, plays a crucial role during cardiovascular development and tumor angiogenesis. To understand its spatiotemporal activity in health and disease is fundamental for the development of drugs to manipulate its activation state. T...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-61781-3 |
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| author | Lukas Herdt Hannes Schihada Michael Kurz Sebastian Ernst Jean Eberlein Peter Kolb Cornelius Krasel Moritz Bünemann Christian S. M. Helker |
| author_facet | Lukas Herdt Hannes Schihada Michael Kurz Sebastian Ernst Jean Eberlein Peter Kolb Cornelius Krasel Moritz Bünemann Christian S. M. Helker |
| author_sort | Lukas Herdt |
| collection | DOAJ |
| description | Abstract The Apelin receptor (APLNR), a class A G-protein coupled receptor, plays a crucial role during cardiovascular development and tumor angiogenesis. To understand its spatiotemporal activity in health and disease is fundamental for the development of drugs to manipulate its activation state. To obtain this understanding, here we develop a tool box of various APLNR conformation biosensors, based on FRET, BRET and the conformation-sensitive fluorophore circularly permuted GFP (cpGFP), with further focus on its in vivo application. We demonstrate the functionality of our biosensors by pharmacological characterization and signal transduction analysis in vitro. Two APLNR-cpGFP biosensors show superior signal-to-noise ratio and are further analyzed for their in vivo applicability. In zebrafish embryos, APLNR-cpGFP biosensors are able to bind both endogenous ligands, Apelin and Apela, and visualize endogenous Aplnr activity in growing blood vessels. Moreover, we are able to measure an Apelin ligand gradient across cellular distances in vivo. Hence, these APLNR conformation biosensors are powerful tools to resolve the spatiotemporal Apelin signaling activity in health and disease. |
| format | Article |
| id | doaj-art-2c449082f97e4f508005ec2b9812988f |
| institution | DOAJ |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-2c449082f97e4f508005ec2b9812988f2025-08-20T03:05:06ZengNature PortfolioNature Communications2041-17232025-07-0116111510.1038/s41467-025-61781-3In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensorLukas Herdt0Hannes Schihada1Michael Kurz2Sebastian Ernst3Jean Eberlein4Peter Kolb5Cornelius Krasel6Moritz Bünemann7Christian S. M. Helker8Department of Biology, Animal Cell Biology, Marburg UniversityInstitute of Pharmaceutical Chemistry, Faculty of Pharmacy, Marburg UniversityInstitute of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Marburg UniversityInstitute of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Marburg UniversityDepartment of Biology, Animal Cell Biology, Marburg UniversityInstitute of Pharmaceutical Chemistry, Faculty of Pharmacy, Marburg UniversityInstitute of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Marburg UniversityInstitute of Pharmacology and Clinical Pharmacy, Faculty of Pharmacy, Marburg UniversityDepartment of Biology, Animal Cell Biology, Marburg UniversityAbstract The Apelin receptor (APLNR), a class A G-protein coupled receptor, plays a crucial role during cardiovascular development and tumor angiogenesis. To understand its spatiotemporal activity in health and disease is fundamental for the development of drugs to manipulate its activation state. To obtain this understanding, here we develop a tool box of various APLNR conformation biosensors, based on FRET, BRET and the conformation-sensitive fluorophore circularly permuted GFP (cpGFP), with further focus on its in vivo application. We demonstrate the functionality of our biosensors by pharmacological characterization and signal transduction analysis in vitro. Two APLNR-cpGFP biosensors show superior signal-to-noise ratio and are further analyzed for their in vivo applicability. In zebrafish embryos, APLNR-cpGFP biosensors are able to bind both endogenous ligands, Apelin and Apela, and visualize endogenous Aplnr activity in growing blood vessels. Moreover, we are able to measure an Apelin ligand gradient across cellular distances in vivo. Hence, these APLNR conformation biosensors are powerful tools to resolve the spatiotemporal Apelin signaling activity in health and disease.https://doi.org/10.1038/s41467-025-61781-3 |
| spellingShingle | Lukas Herdt Hannes Schihada Michael Kurz Sebastian Ernst Jean Eberlein Peter Kolb Cornelius Krasel Moritz Bünemann Christian S. M. Helker In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor Nature Communications |
| title | In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor |
| title_full | In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor |
| title_fullStr | In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor |
| title_full_unstemmed | In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor |
| title_short | In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor |
| title_sort | in vivo measurement of an apelin gradient with a genetically encoded aplnr conformation biosensor |
| url | https://doi.org/10.1038/s41467-025-61781-3 |
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