Topical application of indicators for calcium imaging at the Drosophila larval neuromuscular junction.
Topical application of indicators for calcium imaging at the Drosophila larval neuromuscular junction.
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DOI：
10.1101/pdb.prot070086
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发表时间：
2012-07-01
影响因子：
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通讯作者：
Macleod, Gregory T
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文献类型：
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作者：
Macleod, Gregory T
钙成像是一种技术，在这种技术中，钙结合分子被装载到活细胞中，当它们结合钙(2+)时，它们通过发出的光(荧光或生物发光)的强度或波长的变化来“指示”游离钙的浓度。有几种可能的方法将合成的Ca(2+)指示剂加载到亚细胞室中，包括局部应用膜透入的Ca(2+)指示剂、前向填充葡聚糖结合物和直接注射。钙离子成像是神经生物学中一种信息量很大的技术，因为钙离子参与了许多神经信号通路，并触发了神经递质的释放。本文介绍了Ca(2+)指示剂在果蝇幼虫神经肌肉接头(NMJ)的局部应用。这种加载技术执行起来很简单，并且可以快速生成数据。缺点是数据可能很难解释，主要是因为很难确定哪个细胞和亚细胞隔室(S)被加载(例如，肌肉、神经或胶质细胞；胞浆、线粒体或内质网)。
Calcium imaging is a technique in which Ca(2+)-binding molecules are loaded into live cells and as they bind Ca(2+) they "indicate" the concentration of free calcium through a change in either the intensity or the wavelength of light emitted (fluorescence or bioluminescence). There are several possible methods for loading synthetic Ca(2+) indicators into subcellular compartments, including topical application of membrane-permeant Ca(2+) indicators, forward-filling of dextran conjugates, and direct injection. Calcium imaging is a highly informative technique in neurobiology because Ca(2+) is involved in many neuronal signaling pathways and serves as the trigger for neurotransmitter release. This article describes the topical application of Ca(2+) indicators at the Drosophila larval neuromuscular junction (NMJ). This loading technique is simple to execute and yields data quickly. The drawback is that the data can be difficult to interpret, primarily because it is difficult to ascertain which cellular and subcellular compartment(s) are loaded (e.g., muscle, nerve, or glia; cytosol, mitochondrion, or endoplasmic reticulum).