NDLI: Laser-scanning photostimulation of optogenetically targeted forebrain circuits.

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Year: 2013 Issue: 82

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Laser-scanning photostimulation of optogenetically targeted forebrain circuits.

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Laser-scanning photostimulation of optogenetically targeted forebrain circuits.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Lee, Charles C. Lam, Ying-Wan Imaizumi, Kazuo Sherman, S. Murray
Description	Author Affiliation: Lee CC ( Department of Comparative Biomedical Sciences, School of Veterinary Medicine, Louisiana State University.)
Abstract	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.
File Format	HTM / HTML
e-ISSN	1940087X
DOI	10.3791/50915
Journal	Journal of Visualized Experiments
Issue Number	82
Language	English
Publisher	MyJove Corp.
Publisher Date	2013-12-27
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Physical Sciences Discipline Life Sciences Discipline Medicine Nerve Net Physiology Photic Stimulation Prosencephalon Adenoviridae Genetics Animals Bacterial Proteins Biosynthesis Chemistry Luminescent Proteins Mice Mice, Inbred Balb C Mice, Transgenic Microscopy, Confocal Neurons Recombinant Fusion Proteins Rhodopsin Synapses Transfection Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't Research Support, U.s. Gov't, Non-p.h.s. Video-audio Media
Content Type	Text
Resource Type	Article

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