Simultaneous visualization of multiple neuronal properties with single-cell resolution in the living rodent brain

To understand the fine-scale structures and functional properties of individual neurons in vivo, we developed and validated a rapid genetic technique that enables simultaneous investigation of multiple neuronal properties with single-cell resolution in the living rodent brain. Our technique PASME (p...

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Bibliographic Details
Published in:Molecular and cellular neuroscience 2011-11, Vol.48 (3), p.246-257
Main Authors: Ako, Rie, Wakimoto, Mayu, Ebisu, Haruka, Tanno, Kaori, Hira, Riichiro, Kasai, Haruo, Matsuzaki, Masanori, Kawasaki, Hiroshi
Format: Article
Language:English
Subjects:
Animals
Cell Count
Cerebral cortex
Cerebral Cortex - cytology
Cerebral Cortex - physiology
Electroporation - methods
Gene Transfer Techniques
Hippocampus
In utero electroporation
Mice
Mice, Inbred ICR
Microscopy, Confocal - methods
Multiple-gene labeling
Neurons - cytology
Neurons - physiology
Rats
Rats, Wistar
Sparse-neuron labeling in vivo
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Summary:To understand the fine-scale structures and functional properties of individual neurons in vivo, we developed and validated a rapid genetic technique that enables simultaneous investigation of multiple neuronal properties with single-cell resolution in the living rodent brain. Our technique PASME (promoter-assisted sparse-neuron multiple-gene labeling using in uteroelectroporation) targets specific small subsets of sparse pyramidal neurons in layer 2/3, layer 5 of the cerebral cortex and in the hippocampus with multiple fluorescent reporter proteins such as postsynaptic PSD-95-GFP and GFP-gephyrin. The technique is also applicable for targeting independently individual neurons and their presynaptic inputs derived from surrounding neurons. Targeting sparse layer 2/3 neurons, we uncovered a novel subpopulation of layer 2/3 neurons in the mouse cerebral cortex. This technique, broadly applicable for probing and manipulating neurons with single-cell resolution in vivo, should provide a robust means to uncover the basic mechanisms employed by the brain, especially when combined with in vivo two-photon laser-scanning microscopy and/or optogenetic technologies.
ISSN:1044-7431
1095-9327
DOI:10.1016/j.mcn.2011.08.005