Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System.

Dye Electroporation and Imaging of Calcium Signaling in Xenopus Nervous System.
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非洲爪蟾神经系统钙信号的染料电穿孔和成像

DOI:
10.1007/978-1-4939-8784-9_15
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发表时间:
2018
影响因子:
--
通讯作者:
Manzini I
Manzini I
中科院分区:
--
文献类型:
--
作者:
Weiss L;Offner T;Hassenklöver T;Manzini I

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电穿孔是一种将带电大分子转移到活细胞中以研究其形态、功能和神经网络内连通性的有效方法。用荧光基团偶联的大分子标记细胞可以用来追踪整个神经元群的投影,以及单个细胞的精细形态。在这里,我们提出了一种方案,以非洲爪蟾的嗅觉系统为模型,可视化大脑中感觉接力突触的突触前和突触后成分。我们使用大块电穿孔来追踪从鼻子到大脑的受体神经元的投影,并使用单细胞电穿孔来可视化它们的突触靶细胞,二尖瓣簇细胞的形态。用钙敏感染料标记受体神经元使我们能够通过功能性钙成像记录刺激诱导的突触前输入到突触后细胞的树突。
Electroporation is an efficient method of transferring charged macromolecules into living cells in order to study their morphology, function, and connectivity within neuronal networks. Labeling cells with fluorophore-coupled macromolecules can be used to trace projections of whole neuronal ensembles, as well as the fine morphology of single cells. Here, we present a protocol to visualize pre- and postsynaptic components of a sensory relay synapse in the brain, using the olfactory system ofXenopus laevistadpoles as a model. We apply bulk electroporation to trace projections of receptor neurons from the nose to the brain, and single cell electroporation to visualize the morphology of their synaptic target cells, the mitral-tufted cells. Labeling the receptor neurons with a calcium-sensitive dye allows us to record stimulus-induced presynaptic input to the dendrites of the postsynaptic cells via functional calcium imaging.
爪蟾ta t的嗅觉受体神经元的类别和狭窄的选择性。
DOI: 10.1085/jgp.200308970
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发表时间: 2014-10-30
期刊: Journal of visualized experiments : JoVE
影响因子: --
作者:
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