A new probe for super-resolution imaging of membranes elucidates trafficking pathways.
A new probe for super-resolution imaging of membranes elucidates trafficking pathways.
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DOI：
10.1083/jcb.201402066
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发表时间：
2014-05-26
期刊：
影响因子：
--
通讯作者：
Rizzoli SO
中科院分区：
文献类型：
--
作者：
Revelo NH;Kamin D;Truckenbrodt S;Wong AB;Reuter-Jessen K;Reisinger E;Moser T;Rizzoli SO
mling是一种新型的膜探针，用于研究膜运输，在广泛的生物系统中的活细胞和固定细胞中都有应用价值。由于缺乏合适的标记工具，参与膜回收的细胞器的分子组成难以确定。本文介绍了一种新型探针，称为膜结合荧光团半胱氨酸-赖氨酸-棕榈酰基团（mling），它在质膜上标记，并在胞吞过程中被吸收。它在固定和渗透后仍然附着在膜上，因此可以与免疫染色和超分辨率显微镜结合使用。我们将mcing应用于哺乳动物培养的细胞、酵母、细菌、原代培养的神经元、黑腹果蝇幼虫的神经肌肉连接和哺乳动物组织。mcing使我们能够研究不同运输细胞器的分子组成。我们用它来解决与来自Corti器官的听觉内毛细胞突触囊泡循环有关的几个问题，并研究在培养神经元中主动或自发循环的突触囊泡之间的分子差异。我们得出的结论是，mling可以在广泛的制剂中研究运输膜。
mCLING is a novel membrane probe for the study of membrane trafficking with demonstrated value in both live and fixed cells across a wide range of biological systems. The molecular composition of the organelles involved in membrane recycling is difficult to establish as a result of the absence of suitable labeling tools. We introduce in this paper a novel probe, named membrane-binding fluorophore-cysteine-lysine-palmitoyl group (mCLING), which labels the plasma membrane and is taken up during endocytosis. It remains attached to membranes after fixation and permeabilization and can therefore be used in combination with immunostaining and super-resolution microscopy. We applied mCLING to mammalian-cultured cells, yeast, bacteria, primary cultured neurons, Drosophila melanogaster larval neuromuscular junctions, and mammalian tissue. mCLING enabled us to study the molecular composition of different trafficking organelles. We used it to address several questions related to synaptic vesicle recycling in the auditory inner hair cells from the organ of Corti and to investigate molecular differences between synaptic vesicles that recycle actively or spontaneously in cultured neurons. We conclude that mCLING enables the investigation of trafficking membranes in a broad range of preparations.
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影响因子：
25
作者：
Fredj, Naila Ben;Burrone, Juan
通讯作者：
Burrone, Juan
影响因子：
16.2
作者：
Frank, Thomas;Rutherford, Mark A.;Strenzke, Nicola;Neef, Andreas;Pangrsic, Tina;Khimich, Darina;Fetjova, Anna;Gundelfinger, Eckart D.;Liberman, M. Charles;Harke, Benjamin;Bryan, Keith E.;Lee, Amy;Egner, Alexander;Riedel, Dietmar;Moser, Tobias
通讯作者：
Moser, Tobias
影响因子：
16.2
作者：
Dani, Adish;Huang, Bo;Bergan, Joseph;Dulac, Catherine;Zhuang, Xiaowei
通讯作者：
Zhuang, Xiaowei
影响因子：
14.8
作者：
Beaudoin, Gerard M. J., III;Lee, Seung-Hye;Arikkath, Jyothi
通讯作者：
Arikkath, Jyothi
影响因子：
2.9
作者：
BANKER, GA;COWAN, WM
通讯作者：
COWAN, WM