用基因编码的applnr构象生物传感器在体内测量Apelin梯度

IF 15.7 1区 综合性期刊 Q1 MULTIDISCIPLINARY SCIENCES
Lukas Herdt, Hannes Schihada, Michael Kurz, Sebastian Ernst, Jean Eberlein, Peter Kolb, Cornelius Krasel, Moritz Bünemann, Christian S. M. Helker
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引用次数: 0

摘要

Apelin受体(applnr)是一种a类g蛋白偶联受体,在心血管发育和肿瘤血管生成中起着至关重要的作用。了解其在健康和疾病中的时空活动是开发操纵其激活状态的药物的基础。为了了解这一点,我们基于FRET、BRET和构象敏感的荧光团循环排列GFP (cpGFP),开发了一个包含各种APLNR构象生物传感器的工具箱,并进一步关注其在体内的应用。我们通过体外药理表征和信号转导分析证明了我们的生物传感器的功能。两种APLNR-cpGFP生物传感器表现出优越的信噪比,并进一步分析了它们在体内的适用性。在斑马鱼胚胎中,Aplnr - cpgfp生物传感器能够结合内源性配体Apelin和Apela,并在生长的血管中可视化内源性Aplnr活性。此外,我们能够在体内测量跨细胞距离的Apelin配体梯度。因此,这些applnr构象生物传感器是研究Apelin信号在健康和疾病中的时空活动的有力工具。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor

In vivo measurement of an Apelin gradient with a genetically encoded APLNR conformation biosensor

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.

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来源期刊
Nature Communications
Nature Communications Biological Science Disciplines-
CiteScore
24.90
自引率
2.40%
发文量
6928
审稿时长
3.7 months
期刊介绍: Nature Communications, an open-access journal, publishes high-quality research spanning all areas of the natural sciences. Papers featured in the journal showcase significant advances relevant to specialists in each respective field. With a 2-year impact factor of 16.6 (2022) and a median time of 8 days from submission to the first editorial decision, Nature Communications is committed to rapid dissemination of research findings. As a multidisciplinary journal, it welcomes contributions from biological, health, physical, chemical, Earth, social, mathematical, applied, and engineering sciences, aiming to highlight important breakthroughs within each domain.
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