Small fluorescent molecules for monitoring autophagic flux

被引:75
作者
Iwashita, Hidefumi [1 ,2 ]
Sakurai, Hajime Tajima [3 ]
Nagahora, Noriyoshi [2 ]
Ishiyama, Munetaka [1 ]
Shioji, Kosei [2 ]
Sasamoto, Kazumi [1 ]
Okuma, Kentaro [2 ]
Shimizu, Shigeomi [3 ]
Ueno, Yuichiro [1 ]
机构
[1] Dojindo Labs, Tabaru 2025-5,Mashiki Machi, Kumamoto 8612202, Japan
[2] Fukuoka Univ, Fac Sci, Dept Chem, Fukuoka, Japan
[3] Tokyo Med & Dent Univ, Med Res Inst, Dept Pathol Cell Biol, Tokyo, Japan
关键词
autolysosome; autophagosome; autophagy; fluorescent probe; LYSOSOMES; FUSION;
D O I
10.1002/1873-3468.12979
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
070307 [化学生物学]; 071010 [生物化学与分子生物学];
摘要
We have developed two types of fluorescent probes, DALGreen and DAPGreen, for monitoring autophagy, that exhibit fluorescence upon being incorporated into autophagosomes. DALGreen enhances its fluorescence at acidic pH, which is favorable for monitoring late-phase autophagy, whereas DAPGreen remains fluorescent with almost constant brightness during the autophagic process. With these probes that stain autophagosomes as they are being formed, the real-time change of autophagic phenomena of live cells may be traced, which is an advantage over conventional approaches with small molecules that stain mature autophagosomes. The use of both dyes allows monitoring of the membrane dynamics of autophagy in any type of cell without the need for genetic engineering, and therefore, will be useful as a tool to study autophagic phenomena.
引用
收藏
页码:559 / 567
页数:9
相关论文
共 15 条
[1]
Live Cell Imaging of Mitochondrial Autophagy with a Novel Fluorescent Small Molecule [J].
Iwashita, Hidefumi ;
Torii, Satoru ;
Nagahora, Noriyoshi ;
Ishiyama, Munetaka ;
Shioji, Kosei ;
Sasamoto, Kazumi ;
Shimizu, Shigeomi ;
Okuma, Kentaro .
ACS CHEMICAL BIOLOGY, 2017, 12 (10) :2546-2551
[2]
A two-photon fluorescent probe for real-time monitoring of autophagy by ultrasensitive detection of the change in lysosomal polarity [J].
Jiang, Jiacheng ;
Tian, Xiaohe ;
Xu, Changzhi ;
Wang, Shuxin ;
Feng, Yan ;
Chen, Man ;
Yu, Haizhu ;
Zhua, Manzhou ;
Meng, Xiangming .
CHEMICAL COMMUNICATIONS, 2017, 53 (26) :3645-3648
[3]
LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing [J].
Kabeya, Y ;
Mizushima, N ;
Uero, T ;
Yamamoto, A ;
Kirisako, T ;
Noda, T ;
Kominami, E ;
Ohsumi, Y ;
Yoshimori, T .
EMBO JOURNAL, 2000, 19 (21) :5720-5728
[4]
Does bafilomycin A1 block the fusion of autophagosomes with lysosomes? [J].
Klionsky, Daniel J. ;
Elazar, Zvulun ;
Seglen, Per O. ;
Rubinsztein, David C. .
AUTOPHAGY, 2008, 4 (07) :849-850
[5]
Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice [J].
Komatsu, M ;
Waguri, S ;
Ueno, T ;
Iwata, J ;
Murata, S ;
Tanida, I ;
Ezaki, J ;
Mizushima, N ;
Ohsumi, Y ;
Uchiyama, Y ;
Kominami, E ;
Tanaka, K ;
Chiba, T .
JOURNAL OF CELL BIOLOGY, 2005, 169 (03) :425-434
[6]
The role of autophagy during the early neonatal starvation period [J].
Kuma, A ;
Hatano, M ;
Matsui, M ;
Yamamoto, A ;
Nakaya, H ;
Yoshimori, T ;
Ohsumi, Y ;
Tokuhisa, T ;
Mizushima, N .
NATURE, 2004, 432 (7020) :1032-1036
[7]
LC3, an autophagosome marker, can be incorporated into protein aggregates independent of autophagy [J].
Kuma, Akiko ;
Matsui, Makoto ;
Mizushima, Noboru .
AUTOPHAGY, 2007, 3 (04) :323-328
[8]
Autophagy: eat thyself, sustain thyself [J].
Marx, Vivien .
NATURE METHODS, 2015, 12 (12) :1121-1125
[9]
Autophagy fights disease through cellular self-digestion [J].
Mizushima, Noboru ;
Levine, Beth ;
Cuervo, Ana Maria ;
Klionsky, Danial J. .
NATURE, 2008, 451 (7182) :1069-1075
[10]
Methods in Mammalian Autophagy Research [J].
Mizushima, Noboru ;
Yoshimori, Tamotsu ;
Levine, Beth .
CELL, 2010, 140 (03) :313-326