Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling.
Revealing the tissue-level complexity of endogenous glucagon-like peptide-1 receptor expression and signaling.
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DOI：
10.1038/s41467-022-35716-1
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发表时间：
2023-01-18
影响因子：
16.6
通讯作者：
Hodson, David J.
中科院分区：
文献类型：
--
作者：
Ast, Julia;Nasteska, Daniela;Fine, Nicholas H. F.;Nieves, Daniel J.;Koszegi, Zsombor;Lanoiselee, Yann;Cuozzo, Federica;Viloria, Katrina;Bacon, Andrea;Luu, Nguyet T.;Newsome, Philip N.;Calebiro, Davide;Owen, Dylan M.;Broichhagen, Johannes;Hodson, David J.
胰高血糖素样肽-1受体（GLP 1R）是一种B类G蛋白偶联受体（GPCR），参与葡萄糖稳态和食物摄入。GLP1R激动剂（GLP1RA）被广泛用于治疗糖尿病和肥胖症，但可视化GPCR的内源性定位、组织和动力学迄今为止仍然遥不可及。在本研究中，我们产生了小鼠窝藏的酶自标记基因组编辑到内源性Glp1r基因座。我们还合理设计和测试了各种荧光染料，涵盖青色到远红色波长，用于组织中的标记性能。通过结合这些技术，我们表明，内源性GLP1R可以特异性和敏感性检测原发组织使用多种颜色。GLP1R动态的纵向分析揭示了相邻细胞亚群进入信号传导和运输的异质性招募，在GLP1RA类别和双重激动剂之间观察到差异。在纳米水平上，GLP1Rs被发现具有更高的组织，经历GLP1RA依赖的膜扩散。总之，这些结果显示了酶自标记用于内源性蛋白质的可视化和询问的效用，并提供了对原代细胞和组织中的B类GPCR的生物学的洞察。可视化内源性GPCR具有挑战性。在这里，作者产生了具有酶自标记基因组的小鼠，该基因组被编辑到内源性胰高血糖素样肽-1受体基因座中，设计了用于复杂组织中特异性标记的荧光染料，并揭示了内源性B类GPCR的组织水平组织和动力学。
The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in glucose homeostasis and food intake. GLP1R agonists (GLP1RA) are widely used in the treatment of diabetes and obesity, yet visualizing the endogenous localization, organization and dynamics of a GPCR has so far remained out of reach. In the present study, we generate mice harboring an enzyme self-label genome-edited into the endogenous Glp1r locus. We also rationally design and test various fluorescent dyes, spanning cyan to far-red wavelengths, for labeling performance in tissue. By combining these technologies, we show that endogenous GLP1R can be specifically and sensitively detected in primary tissue using multiple colors. Longitudinal analysis of GLP1R dynamics reveals heterogeneous recruitment of neighboring cell subpopulations into signaling and trafficking, with differences observed between GLP1RA classes and dual agonists. At the nanoscopic level, GLP1Rs are found to possess higher organization, undergoing GLP1RA-dependent membrane diffusion. Together, these results show the utility of enzyme self-labels for visualization and interrogation of endogenous proteins, and provide insight into the biology of a class B GPCR in primary cells and tissue. Visualizing endogenous GPCRs is challenging. Here the authors generate mice with an enzyme self-label genome-edited into the endogenous glucagon-like peptide-1 receptor locus, design fluorescent dyes for specific labelling in complex tissue, and reveal tissue-level organisation and dynamics of an endogenous class B GPCR.
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DOI：
10.1073/pnas.2107665118
发表时间：
2021-09-14
影响因子：
11.1
作者：
Kemter E;Müller A;Neukam M;Ivanova A;Klymiuk N;Renner S;Yang K;Broichhagen J;Kurome M;Zakhartchenko V;Kessler B;Knoch KP;Bickle M;Ludwig B;Johnsson K;Lickert H;Kurth T;Wolf E;Solimena M
通讯作者：
Solimena M
影响因子：
16.6
作者：
Jones B;Buenaventura T;Kanda N;Chabosseau P;Owen BM;Scott R;Goldin R;Angkathunyakul N;Corrêa IR Jr;Bosco D;Johnson PR;Piemonti L;Marchetti P;Shapiro AMJ;Cochran BJ;Hanyaloglu AC;Inoue A;Tan T;Rutter GA;Tomas A;Bloom SR
通讯作者：
Bloom SR
影响因子：
10.9
作者：
Benninger, Richard K. P.;Piston, David W.
通讯作者：
Piston, David W.
影响因子：
16.6
作者：
Bevacqua RJ;Dai X;Lam JY;Gu X;Friedlander MSH;Tellez K;Miguel-Escalada I;Bonàs-Guarch S;Atla G;Zhao W;Kim SH;Dominguez AA;Qi LS;Ferrer J;MacDonald PE;Kim SK
通讯作者：
Kim SK
影响因子：
9.8
作者：
Buenaventura, Teresa;Bitsi, Stavroula;Tomas, Alejandra
通讯作者：
Tomas, Alejandra