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Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins
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 |
URI: | http://hdl.handle.net/2115/86630 |
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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