This work was supported by the life insurance company "Novalis-Taitbout", Ecole des Neurosciences de Paris (ENP), the Agence Nationale de la Recherche "ANR-09-NEUR-004" in the frame of "ERA-NET NEURON" of FP7 program by the European Commission, "Foundation pour la Recherche Medicale", the Letten Foundation and the Pasteur Foundation.

1. Stereotaxic injections
<ol>
	<li>The virus used in these experiments is a replication-deficient lentiviral vector based on the HIV virus to express Channelrhodopsine2-YFP (yellow fluorescent protein) fusion construct driven by a synapsin promoter6. The plasmid was generously provided by K. Deisseroth's research group (<a href="http://www.stanford.edu/group/dlab/optogenetics/index.html">http://www.stanford.edu/group/dlab/optogenetics/index.html</a>). Lentiviruses can be obtained from commercial sources o...

<ol>
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No conflicts of interest declared.

Optogenetic tools have recently increased our control over selective neuronal populations, including adult born neurons in the olfactory system. Here we describe how to perform a stereotaxic injection of a lentiviral vector expressing ChR2-YFP in a specific population of adult-born neurons. By carefully monitoring the period post-injection, we are able to analyze and manipulate the activity of a cohort of adult-born neurons with a relative homogeneous age.
Because of their low diffusion spread...

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			<td>Drummond Scientific</td>
			<td>3-000-203-G/X</td>
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			<td>Nanoject II Microinjector</td>
			<td>Drummond Scientific</td>
			<td>3-000-204</td>
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			<td>Fine Science Tools</td>
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</table>

selective viral transduction of adult-born olfactory neurons for chronic in vivo optogenetic stimulation

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells in the walls of the lateral ventricle give rise to neuroblasts that migrate for several millimeters along the rostral migratory stream (RMS) to reach and incorporate into the olfactory bulb. To study the different steps and the impact of adult-born neuron integration into preexisting olfactory circuits, it is necessary to selectively label and manipulate the activity of this specific population of neurons. The recent development of optogenetic technologies offers the opportunity to use light to precisely activate this specific cohort of neurons without affecting surrounding neurons4,5. Here, we present a series of procedures to virally express Channelrhodopsin2(ChR2)-YFP in a temporally restricted cohort of neuroblasts in the RMS before they reach the olfactory bulb and become adult-born neurons. In addition, we show how to implant and calibrate a miniature LED for chronic in vivo stimulation of ChR2-expressing neurons.

Watch this Scientific Journal Video about Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation at JoVE.com

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells ...

selective-viral-transduction-adult-born-olfactory-neurons-for-chronic

Research

JoVE Journal

Neuroscience

15.4K visualizações.  Institut Pasteur and Centre National de la Recherche Scientifique (CNRS). Neurônios adultos-nascido do bulbo olfativo pode ser controlado usando optogenetically Channelrhodopsin2 expressando injeção lentiviral no fluxo migratório rostral e fotoestimulação crônica com uma miniatura implantado LED.