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首页> 外文期刊>The European Journal of Neuroscience >Caged compounds for multichromic optical interrogation of neural systems
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Caged compounds for multichromic optical interrogation of neural systems

机译：用于神经系统的多晶硅光学询问的笼式化合物

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Caged compounds are widely used by neurophysiologists to study many aspects of cellular signaling in glia and neurons. Biologically inert before irradiation, they can be loaded into cells via patch pipette or topically applied in situ to a defined concentration; photolysis releases the caged compound in a very rapid and spatially defined way. As caged compounds are exogenous optical probes, they include not only natural products such neurotransmitters, calcium and IP3 but non-natural products such as fluorophores, drugs and antibodies. In this Technical Spotlight we provide a short introduction to the uncaging technique by discussing the nitroaromatic caging chromophores most widely used in such experiments [e.g. -carboxy-ortho-nitrobenyl (CNB), dimethoxynitrobenzyl (DMNB), 4-methoxy-7-nitroindolinyl (MNI) and 4-carboxymethoxy-7-nitroindolinyl (CDNI)]. We show that recently developed caging chromophores [rutheniumbipyridial (RuBi) and 7-diethylaminocoumarin (DEAC)450] that are photolyzed with blue light (similar to 430-480nm range) can be combined with traditional nitroaromatic caged compounds to enable two-color optical probing of neuronal function. For example, one-photon uncaging of either RuBi-GABA or DEAC450-GABA with a 473-nm laser is facile, and can block nonlinear currents (dendritic spikes or action potentials) evoked by two-photon uncaging of CDNI-Glu at 720nm. We also show that two-photon uncaging of DEAC450-Glu and CDNI-GABA at 900 and 720nm, respectively, can be used to fire and block action potentials. Our experiments illustrate that recently developed chromophores have taken uncaging out of the monochrome era', in which it has existed since 1978, so as to enable multichromic interrogation of neuronal function with single-synapse precision.
机译：神经生理学家广泛使用笼式化合物，以研究胶质胶和神经元细胞信号传导的许多方面。在照射前生物学惰性，它们可以通过贴剂移液管加载到细胞中，或者原位地施加到限定的浓度;光解以非常快速和空间定义的方式释放笼中的化合物。由于笼式化合物是外源性光学探针，它们不仅包括天然产物，如神经递质，钙和IP3，但非天然产品如荧光团，药物和抗体。在本技术聚光灯中，我们通过讨论在此类实验中最广泛使用的硝基芳族母猪发作发色团来提供对未致密技术的简要介绍[例如 - 羧基 - 邻硝基苯基（CNB），二甲氧基苄基（DMNB），4-甲氧基-7-硝基吲哚啉基（MNI）和4-羧甲氧基-7-硝基吲哚啉基（CDNI）]。我们表明，最近开发的CAIRCOOMOCHORS [rutheniumBioIdivial（RUBI）和7-二乙基氨基脲草蛋白（DEAC）450]用蓝光（类似于430-480nm范围），可以与传统的硝基芳族笼式化合物组合，以实现双色光学探测神经元功能。例如，具有473-NM激光的鲁布-GABA或DEAC450-GABA的单光子未展示是容易的，并且可以在720nm处通过双光子未纳入CDNI-Glu诱导的非线性电流（树突状尖峰或动作电位）。我们还表明，分别为900和720nm的DEAC450-GLU和CDNI-GABA的双光子未展示，可用于发射和阻止动作电位。我们的实验说明了最近开发的发色团已经从1978年以来已经存在的单色时代脱出，以便使多晶体函数与单突出精度相比进行多晶体函数。

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