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

被引：1

作者：

Tsuzuki, Atsushi ^{[1
,2
,3
]}

Fujioka, Yoichiro ^{[1
,2
,4
]}

Yoshida, Aiko ^{[1
,2
,4
]}

Kashiwagi, Sayaka ^{[1
,2
,4
]}

Amano, Maho ^{[1
,2
]}

Hira, Tohru ^{[5
]}

Nakamura, Akinobu ^{[2
,3
]}

Miyoshi, Hideaki ^{[2
,3
,6
]}

Atsumi, Tatsuya ^{[2
,3
]}

Ohba, Yusuke ^{[1
,2
,4
,7
]}

机构：

[1] Hokkaido Univ, Fac Med, Dept Cell Physiol, Sapporo, Hokkaido, Japan

[2] Hokkaido Univ, Grad Sch Med, Sapporo, Hokkaido, Japan

[3] Hokkaido Univ, Fac Med, Dept Rheumatol Endocrinol & Nephrol, Sapporo, Hokkaido, Japan

[4] Hokkaido Univ, Global Stn Biosurfaces & Drug Discovery, Sapporo, Hokkaido, Japan

[5] Hokkaido Univ, Div Fundamental AgriSci, Grad Sch Agr, Res Grp Biosci & Chem, Sapporo, Hokkaido, Japan

[6] Hokkaido Univ, Fac Med, Div Diabet & Obes, Sapporo, Hokkaido, Japan

[7] Japan Agcy Med Res & Dev, AMED CREST, Sapporo, Hokkaido, Japan

来源：

JOURNAL OF DIABETES INVESTIGATION | 2022年 / 13卷 / 07期

关键词：

Exocytosis; Glucagon-like peptide-1; Total internal reflection fluorescence microscope; BIOLOGY;

D O I：

10.1111/jdi.13800

中图分类号：

R5 [内科学];

学科分类号：

1002 ; 100201 ;

摘要：

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.

引用

页码：1134 / 1139

页数：6

共 15 条

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