Monitoring lipid droplet dynamics in living cells by using fluorescent probes
Monitoring lipid droplet dynamics in living cells by using fluorescent probes
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使用荧光探针监测活细胞中的脂滴动态
DOI：
10.1021/acs.biochem.8b01071
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发表时间：
2019
期刊：
影响因子：
2.9
通讯作者：
Ueno Y
中科院分区：
文献类型：
--
作者：
Tatenaka Y;Kato H;Ishiyama M;Sasamoto K;Shiga M;Nishitoh H;Ueno Y
我们开发了三种类型的脂滴（LD）特异性荧光探针用于活细胞成像，脂滴-蓝，脂滴-绿和脂滴-红，它们在结合到活细胞和固定细胞的LD时都表现出荧光。这些lipi探针是LD特异性探针，含有芘或苝基团作为荧光支架，可以通过多个细胞器特异性探针同时染色来观察活细胞中的LD动力学以及与其他细胞器的相互关系。此外，Lipi-Blue和Lipi-Green可以在染色后48小时内监测活细胞的ld。在这里，我们发现使用这些探针可以在单个细胞中分别监测新形成的ld和先前存在的ld，并且在相同的培养条件下，在KB细胞中观察到整个ld的细胞间转移，而在HepG2细胞中则没有。这些发现表明，新开发的ld特异性探针可用于分析活细胞中ld的动力学。
We have developed three types of lipid droplet (LD)-specific fluorescent probes for live-cell imaging, Lipi-Blue, Lipi-Green, and Lipi-Red, which exhibit fluorescence upon being incorporated into LDs both of living and of fixed cells. These Lipi-probes are LD-specific probes that contain a pyrene or perylene group as a fluorescent scaffold and can be used to observe dynamics of LD in live cells and also interrelations with other organelles by simultaneous staining with multiple organelle-specific probes. Additionally, Lipi-Blue and Lipi-Green allow monitoring LDs in live cells even for 48 h after the staining. Here we show that newly formed LDs and previously existed LDs can be separately monitored in a single cell by using these probes and that intercellular transfer of whole LDs is observed in KB cells, but not in HepG2 cells under the same culturing condition. These findings indicate that newly developed LD-specific probes are useful to analyze the dynamics of LDs in live cells.