Laser-scanning photostimulation of optogenetically targeted forebrain circuits

The sensory forebrain is composed of intricately connected cell types, of which functional properties have yet to be fully elucidated. Understanding the interactions of these forebrain circuits has been aided recently by the development of optogenetic methods for light-mediated modulation of neurona...

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Bibliographic Details
Published in:Journal of visualized experiments 2013-12 (82), p.50915-50915
Main Authors: Lee, Charles C, Lam, Ying-Wan, Imaizumi, Kazuo, Sherman, S Murray
Format: Article
Language:English
Subjects:
Adenoviridae - genetics
Animals
Bacterial Proteins - biosynthesis
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Luminescent Proteins - biosynthesis
Luminescent Proteins - chemistry
Luminescent Proteins - genetics
Mice
Mice, Inbred BALB C
Mice, Transgenic
Microscopy, Confocal
Nerve Net - physiology
Neurons - physiology
Neuroscience
Photic Stimulation - methods
Prosencephalon - physiology
Recombinant Fusion Proteins - biosynthesis
Recombinant Fusion Proteins - chemistry
Recombinant Fusion Proteins - genetics
Rhodopsin - biosynthesis
Rhodopsin - chemistry
Rhodopsin - genetics
Synapses - physiology
Transfection
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Summary:The sensory forebrain is composed of intricately connected cell types, of which functional properties have yet to be fully elucidated. Understanding the interactions of these forebrain circuits has been aided recently by the development of optogenetic methods for light-mediated modulation of neuronal activity. Here, we describe a protocol for examining the functional organization of forebrain circuits in vitro using laser-scanning photostimulation of channelrhodopsin, expressed optogenetically via viral-mediated transfection. This approach also exploits the utility of cre-lox recombination in transgenic mice to target expression in specific neuronal cell types. Following transfection, neurons are physiologically recorded in slice preparations using whole-cell patch clamp to measure their evoked responses to laser-scanning photostimulation of channelrhodopsin expressing fibers. This approach enables an assessment of functional topography and synaptic properties. Morphological correlates can be obtained by imaging the neuroanatomical expression of channelrhodopsin expressing fibers using confocal microscopy of the live slice or post-fixed tissue. These methods enable functional investigations of forebrain circuits that expand upon more conventional approaches.
ISSN:1940-087X
1940-087X
DOI:10.3791/50915