Imaging calcium dynamics in the nervous system by means of ballistic delivery of indicators
Imaging calcium dynamics in the nervous system by means of ballistic delivery of indicators
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DOI：
10.1016/s0165-0270(02)00154-1
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发表时间：
2002-09-15
影响因子：
3
通讯作者：
Gan, WB
中科院分区：
文献类型：
--
作者：
Kettunen, P;Demas, J;Gan, WB
荧光钙指示剂的使用，超过。多年来，解开了神经元功能和发育的重要钙依赖机制。然而，与钙指示剂的加载相关的困难限制了它们的广泛使用，特别是对于成人神经系统中神经元加工的研究。在这里，我们表明，在中枢和外周神经系统中，神经元及其过程，包括树突棘和丝状伪足的群体，可以快速有效地通过使用“基因枪”提供钙指示剂涂层颗粒标记。重要的是，神经元标记发生在体外和体内，并在广泛的年龄和准备。标记的细胞表现出自发和诱发的钙瞬变，表明粒子介导的传递是无害的神经元功能。此外，与使用贴片移液管加载不同，在弹道加载后保留了细胞质内容物。这使得钙依赖性第二信使途径的研究没有损失的信号成分。因此，钙指示剂的弹道递送为进一步探索神经系统的结构和功能开辟了许多新的途径，从单个棘到神经元网络。(C)2002 Elsevier Science B.V.保留所有权利。
The use of fluorescence-based calcium indicators has, over. the years, unraveled important calcium-dependent mechanisms underlying neuronal function and development. However, difficulties associated with the loading of calcium indicators have limited their widespread use, particularly for the study of neuronal processing in the adult nervous system. Here, we show that in the central and peripheral nervous systems, populations of neurons and their processes, including dendritic spines and filopodia, can be labeled rapidly and efficiently by delivering calcium indicator-coated particles using a 'gene gun'. Importantly, neuronal labeling occurred both in vitro and in vivo, and across a wide range of ages and preparations. The labeled cells demonstrate spontaneous and evoked calcium transients, indicating that particle-mediated delivery is not deleterious to neuronal function. Furthermore, unlike loading with patch pipettes, cytoplasmic content is preserved following ballistic loading. This enables the study of calcium-dependent second messenger pathways without loss of signaling components. The ballistic delivery of calcium indicators thus opens up many new avenues for further exploration of the structure and function of the nervous system from single spines to neuronal networks. (C) 2002 Elsevier Science B.V. All rights reserved.