Whole-Brain Calcium Imaging in Larval Xenopus.

Whole-Brain Calcium Imaging in Larval Xenopus.
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非洲爪蟾幼虫的全脑钙成像

DOI:
10.1101/pdb.prot106815
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发表时间:
2020
影响因子:
--
通讯作者:
Manzini I
Manzini I
中科院分区:
--
文献类型:
--
作者:
Offner T;Daume D Weiss L;Hassenklöver T;Manzini I

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感觉系统检测环境刺激并将其转换为可由中枢神经系统解释的电活动模式。最初的感觉输入在到达更高级的大脑中心的过程中,被插入的次级处理中心连续地转换。绘制所有这些阶段的神经元活动模式对于理解感觉信息处理至关重要。非洲爪蟾幼虫非常适合神经元活动的全脑成像。这主要是由于其体积小,透明,以及感觉系统的外围和中央部分的可访问性。在这里,我们描述了一个协议的钙成像在几个层次的嗅觉系统使用化学钙指示染料或非洲爪蟾转基因系神经元GCaMP6s表达的局部注射。结合快速体积多光子显微镜,所描述的钙成像方法可以提供详细的洞察整个大脑区域的时空活动在不同阶段的感觉信息处理。虽然这些方法广泛适用于中枢神经系统,但在这项工作中,我们专注于嗅球中肾小球和嗅杏仁核中气味反应神经元的钙成像协议。
Sensory systems detect environmental stimuli and transform them into electrical activity patterns interpretable by the central nervous system. En route to higher brain centers, the initial sensory input is successively transformed by interposed secondary processing centers. Mapping the neuronal activity patterns at all of those stages is essential to understand sensory information processing. Larval Xenopus laevis is very well-suited for whole-brain imaging of neuronal activity. This is mainly due to its small size, transparency, and the accessibility of both peripheral and central parts of sensory systems. Here we describe a protocol for calcium imaging at several levels of the olfactory system using focal injection of chemical calcium indicator dyes or a Xenopus transgenic line with neuronal GCaMP6s expression. In combination with fast volumetric multiphoton microscopy, the calcium imaging methods described can provide detailed insight into spatiotemporal activity of entire brain regions at different stages of sensory information processing. Although the methods are broadly applicable to the central nervous system, in this work we focus on protocols for calcium imaging of glomeruli in the olfactory bulb and odor-responsive neurons in the olfactory amygdala.
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