Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics.,Nature Communications

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Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics.
Nature Communications ( IF 14.7 ) Pub Date : 2020-01-24 , DOI: 10.1038/s41467-020-14309-w
Julia Ast _{1,

2} , Anastasia Arvaniti _{1,

2} , Nicholas H F Fine _{1,

2} , Daniela Nasteska _{1,

2} , Fiona B Ashford _{1,

2} , Zania Stamataki ₃ , Zsombor Koszegi _{1,

2} , Andrea Bacon ₄ , Ben J Jones ₅ , Maria A Lucey ₅ , Shugo Sasaki ₆ , Daniel I Brierley ₇ , Benoit Hastoy ₈ , Alejandra Tomas ₉ , Giuseppe D'Agostino ₁₀ , Frank Reimann ₁₁ , Francis C Lynn ₆ , Christopher A Reissaus ₁₂ , Amelia K Linnemann ₁₂ , Elisa D'Este ₁₃ , Davide Calebiro _{1,

2} , Stefan Trapp ₇ , Kai Johnsson ₁₄ , Tom Podewin ₁₄ , Johannes Broichhagen ₁₄ , David J Hodson _{1,

2}

Affiliation

Institute of Metabolism and Systems Research (IMSR), and Centre of Membrane Proteins and Receptors (COMPARE), University of Birmingham, Birmingham, UK.
Centre for Endocrinology, Diabetes and Metabolism, Birmingham Health Partners, Birmingham, UK.
Centre for Liver Research, College of Medical and Dental Sciences, Institute for Immunology and Immunotherapy, University of Birmingham, Birmingham, UK.
Genome Editing Facility, Technology Hub, University of Birmingham, Birmingham, UK.
Division of Diabetes, Endocrinology and Metabolism, Section of Investigative Medicine, Imperial College London, London, UK.
Diabetes Research Group, BC Children's Hospital Research Institute, Vancouver, BC, Canada; Department of Surgery, University of British Columbia, Vancouver, BC, Canada.
Centre for Cardiovascular and Metabolic Neuroscience, Department of Neuroscience, Physiology & Pharmacology, University College London, London, UK.
Oxford Centre for Diabetes, Endocrinology & Metabolism, University of Oxford, Oxford, UK.
Division of Diabetes, Endocrinology and Metabolism, Section of Cell Biology and Functional Genomics, Imperial College London, London, UK.
Faculty of Biology, Medicine and Health, University of Manchester, Oxford Road, Manchester, UK.
Wellcome Trust-MRC Institute of Metabolic Science, University of Cambridge, Cambridge, UK.
Department of Pediatrics, Indiana University School of Medicine, Indianapolis, IN, USA.
Optical Microscopy Facility, Max Planck Institute for Medical Research, Heidelberg, Germany.
Department of Chemical Biology, Max Planck Institute for Medical Research, Heidelberg, Germany.

The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in metabolism. Presently, its visualization is limited to genetic manipulation, antibody detection or the use of probes that stimulate receptor activation. Herein, we present LUXendin645, a far-red fluorescent GLP1R antagonistic peptide label. LUXendin645 produces intense and specific membrane labeling throughout live and fixed tissue. GLP1R signaling can additionally be evoked when the receptor is allosterically modulated in the presence of LUXendin645. Using LUXendin645 and LUXendin651, we describe islet, brain and hESC-derived β-like cell GLP1R expression patterns, reveal higher-order GLP1R organization including membrane nanodomains, and track single receptor subpopulations. We furthermore show that the LUXendin backbone can be optimized for intravital two-photon imaging by installing a red fluorophore. Thus, our super-resolution compatible labeling probes allow visualization of endogenous GLP1R, and provide insight into class B GPCR distribution and dynamics both in vitro and in vivo.

中文翻译：

超分辨率显微镜兼容荧光探针揭示了内源性胰高血糖素样肽 1 受体分布和动力学。

胰高血糖素样肽 1 受体 (GLP1R) 是一类参与代谢的 BG 蛋白偶联受体 (GPCR)。目前，其可视化仅限于基因操作、抗体检测或使用刺激受体激活的探针。在这里，我们提出了 LUXendin645，一种远红荧光 GLP1R 拮抗肽标签。LUXendin645 在整个活组织和固定组织中产生强烈和特异性的膜标记。当受体在 LUXendin645 存在的情况下进行变构调节时，还可以诱发 GLP1R 信号传导。使用 LUXendin645 和 LUXendin651，我们描述了胰岛、脑和 hESC 衍生的 β 样细胞 GLP1R 表达模式，揭示了包括膜纳米结构域在内的高阶 GLP1R 组织，并跟踪单个受体亚群。我们进一步表明，通过安装红色荧光团，可以优化 LUXendin 骨架以用于活体双光子成像。因此，我们的超分辨率兼容标记探针允许内源性 GLP1R 的可视化，并提供对 B 类 GPCR 分布和体外和体内动力学的洞察。

更新日期：2020-01-24

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