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Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins

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Title: Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins
Authors: Tsuzuki, Atsushi Browse this author
Fujioka, Yoichiro Browse this author →KAKEN DB
Yoshida, Aiko Browse this author →KAKEN DB
Kashiwagi, Sayaka Browse this author →KAKEN DB
Amano, Maho Browse this author →KAKEN DB
Hira, Tohru Browse this author →KAKEN DB
Nakamura, Akinobu Browse this author →KAKEN DB
Miyoshi, Hideaki Browse this author →KAKEN DB
Atsumi, Tatsuya Browse this author →KAKEN DB
Ohba, Yusuke Browse this author →KAKEN DB
Keywords: Exocytosis
Glucagon-like peptide-1
Total internal reflection fluorescence microscope
Issue Date: Jul-2022
Publisher: John Wiley & Sons
Journal Title: Journal of diabetes investigation
Volume: 13
Issue: 7
Start Page: 1134
End Page: 1139
Publisher DOI: 10.1111/jdi.13800
Abstract: Live-cell imaging with fluorescent proteins (FPs) is a powerful tool for investigating the exocytosis processes of hormones. However, the secretion process of glucagon-like peptide-1 (GLP-1) has not been visualized by FPs, which might be because tagging FPs inhibits GLP-1 synthesis through the post-translational processing from proglucagon. Here, we have developed FP-tagged GLP-1 by inserting FPs into the middle of GLP-1 and adding the proglucagon signal peptide. Confocal imaging confirmed that GLP-1 fused to FPs with high folding efficiency showed granular structure, in which secretory vesicle markers colocalized. The fluorescence intensity of FP in the culture supernatant from cells treated with KCl or forskolin was significantly increased compared with those from untreated cells. Furthermore, FP-tagged GLP-1 enables direct visualization of stimulation-dependent exocytosis of GLP-1 at a single granule resolution with total internal reflection fluorescence microscopy. FP-tagged GLP-1 might facilitate the screening of GLP-1 secretagogues and the discovery of new antidiabetic drugs.
Type: article
Appears in Collections:国際連携研究教育局 : GI-CoRE (Global Institution for Collaborative Research and Education : GI-CoRE) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)
医学院・医学研究院 (Graduate School of Medicine / Faculty of Medicine) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

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