Name: selective viral transduction of adult-born olfactory neurons for chronic in vivo optogenetic stimulation
Uploaded: 2011-12-28T12:00:00
Description: Watch this Scientific Journal Video about Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation at JoVE.com

Diese Arbeit wurde von der Versicherungsgesellschaft &quot;Novalis-Taitbout&quot;, Ecole des Neurosciences de Paris (ENP), die Agence Nationale de la Recherche &quot;ANR-09-neur-004&quot; im Rahmen des &quot;ERA-NET NEURON&quot; der unterstützten RP7-Programm von der Europäischen Kommission, &quot;Foundation pour la Recherche Medicale&quot;, die Letten-Stiftung und der Pasteur-Stiftung.

1. Stereotaktische Injektion <ol><li> Das Virus in diesen Experimenten verwendet wird ein Replikations-defizienten lentiviralen Vektors auf das HIV-Virus basiert auf Channelrhodopsine2-YFP (gelb fluoreszierendes Protein) exprimieren Fusionskonstrukt angetrieben von einem Synapsin Förderer 6. Das Plasmid wurde freundlicherweise von K. Deisseroth &#39;s Research Group (sofern <a href="http://www.stanford.edu/group/dlab/optogenetics/index.html">http://www.stanford.edu/group/dlab/optogenetics/index.html</a> )....

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Optogenetische Werkzeuge haben vor kurzem unsere Kontrolle über selektive neuronale Populationen, einschließlich der Erwachsenenbildung geboren Neuronen im olfaktorischen System erhöht. Hier beschreiben wir, wie Sie eine stereotaktische Injektion eines lentiviralen Vektor, ChR2-YFP in einer bestimmten Population von Erwachsenen geboren Neuronen führen. Durch die sorgfältige Überwachung der Zeit nach der Injektion, sind wir in der Lage zu analysieren und zu manipulieren, die Aktivität von einer Kohorte von Erwachs...

<table border="1"><tbody><tr><td> Material </td><td> Firma </td><td> Katalog Nr. </td></tr><tr><td> Ketamin </td><td> Merial </td><td> Imalgène 1000 </td></tr><tr><td> Xylazin </td><td> Bayer Health Care </td><td> Rompum 0,2% </td></tr><tr><td> Pipette Puller </td><td> Sutter </td><td> P-97 </td></tr><tr><td> Glaskapillaren (3,5 &quot;) </td><td> Drummond Scientific </td><td> 3-000-203-G / X </td></tr><tr><td> Nanoject II Mikroinjektor </td><td> Drummond Scientific </td><td> 3-000-204 </td></tr><tr><td> High-Speed-chirurgische Bohrer </td><td> Feine Science Tools </td><td> 18000-17 </td></tr><tr><td> Micro Drill Stahl Grate 0,7 mm Spitzendurchmesser </td><td> Feine Science Tools </td><td> 19008-07 </td></tr><tr><td> LED CMS blau 4,6 W </td><td> OSRAM </td><td> LBW5SN </td></tr><tr><td> Kabeldose (2,54 mm) </td><td> Fischer Elektronik </td><td> BL5.36Z </td></tr><tr><td> Stromregler für LED (max: 1A) </td><td> A1W Electronik </td><td> HKO-KL50-1000 </td></tr><tr><td> Impulsgeber </td><td> AMPI </td><td> Meister-8 </td></tr><tr><td> Optisches Leistungsmessgerät </td><td> Thor Labs </td><td> PM100 </td></tr><tr><td> Cyanacrylat-Klebstoff </td><td> Loctite </td><td> 454 </td></tr><tr><td> Polycarboxylatzement (erste Schicht Zement) </td><td> Septodont </td><td> Selfast (# 0068Q) </td></tr><tr><td> Methacrylat-Zement (zweite Schicht Zement) </td><td> GC InternationalCorp. </td><td> Unifast Trad - Liquid (# 339292) Unifast Trad - Pulver (# 339114) </td></tr><tr><td> Carprofen </td><td> Pfizer </td><td> Rimadyl </td></tr></tbody></table>

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

Lokale Interneurone werden kontinuierlich in den Riechkolben der erwachsenen Nagetieren 1-3 regeneriert. In diesem Prozess, genannt adulte Neurogenese, neuronale Stammzellen in den Wänden der Seitenventrikel Anlass zu Neuroblasten, die für mehrere Millimeter entlang der rostralen wandernden Strom (RMS) zu erreichen und in den Riechkolben enthalten migrieren. Zur Untersuchung der verschiedenen Schritte und die Auswirkungen der Erwachsenen-geboren Neuron Integration in bereits vorhandene Geruchs-Schaltungen ist es notwendig, gezielt Label und Manipulation der Aktivität dieser speziellen Population von Neuronen. Die jüngste Entwicklung der optogenetische Technologien bietet die Möglichkeit, Licht zu verwenden, um genau zu aktivieren diese spezifische Gruppe von Neuronen, ohne die umliegenden Neuronen 4,5. Hier präsentieren wir eine Reihe von Verfahren, die viral ausdrückliche Channelrhodopsin2 (ChR2)-YFP in einer zeitlich beschränkten Gruppe von Neuroblasten in der RMS, bevor sie den Riechkolben zu erreichen und zu Erwachsenen-geboren neurons. Darüber hinaus zeigen wir, wie zu implantieren und zu kalibrieren eine Miniatur für chronische in vivo Stimulation von ChR2-exprimierenden Neuronen LED.

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-geboren Neuronen des Bulbus kann optogenetically gesteuert werden mit Channelrhodopsin2-exprimierenden lentiviralen Injektion in den rostralen wandernde Bach und chronischen Photostimulation mit einem implantierten Mini-LED.

Selektive Viral Transduktion von Adult-geboren olfaktorischen Neuronen für chronische In vivo Optogenetische Stimulation

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

Research

JoVE Journal

Neuroscience

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

Studying the Integration of Adult-born Neurons

Die Untersuchung der Integration von Adult-geboren Neurons

Neurogenesis occurs in adult mammalian brains in the sub-ventricular zone (SVZ) of the lateral ventricle and in the sub-granular zone (SGZ) of the ...

Forschung

Neurowissenschaften

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-mediated Genetic Manipulation and Visualization of Olfactory Sensory Neurons in vivo

Lentivirus-vermittelte Gen-Manipulation und Visualisierung von Riechzellen In vivo</em

Development of a precise olfactory circuit relies on accurate projection of olfactory sensory neuron (OSN) axons to their synaptic 
targets in the ...

Flash Photolysis of Caged Compounds in the Cilia of Olfactory Sensory  Neurons

Flash-Photolyse von Caged Compounds in den Zilien der Riechzellen

Photolysis of caged compounds allows the production of rapid and localized increases in the concentration of various physiologically active compounds1. ...

Fiber-optic Implantation for Chronic Optogenetic Stimulation of Brain Tissue

LWL-Implantation für chronische optogenetische Stimulation des Hirngewebes

Elucidating patterns of neuronal connectivity has been a challenge for both clinical and basic neuroscience. Electrophysiology has been the gold standard ...

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetic Stimulation of Escape Behavior in Drosophila melanogaster

Optogenetische Stimulation von Escape Verhalten in<em&gt; Drosophila melanogaster</em

A  growing number of genetically encoded tools are becoming available that allow non-invasive  manipulation of the neural activity of specific neurons in ...

A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo

A Method for High Fidelity Optogenetic Control of Individual Pyramidal Neurons In vivo

Eine Methode zur High Fidelity optogenetische Kontrolle der Einzelpyramidenzellen<em&gt; In-vivo-</em

Optogenetic methods  have emerged as a powerful tool for elucidating neural circuit activity  underlying a diverse set of behaviors across a broad range ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo optogenetische Stimulation des Nager Central Nervous System

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

Intracerebroventricular Viral Injection of the Neonatal Mouse Brain for Persistent and Widespread Neuronal Transduction

Intracerebroventricular Viral Injektion der Neonatal Maus Gehirn für anhaltenden und verbreiteten Neuronale Transduction

With the pace of scientific advancement accelerating rapidly, new methods are needed for experimental neuroscience to quickly and easily manipulate gene ...

Optogenetic Stimulation of the Auditory Nerve

Optogenetische Stimulation des Hörnervs

Direct electrical stimulation of spiral ganglion neurons (SGNs) by cochlear implants (CIs) enables open speech comprehension in the majority of implanted ...

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Ex Vivo Optogenetic Dissection of Fear Circuits in Brain Slices

Ex Vivo optogenetische Dissection of Fear Schaltkreise in Hirnschnitten

Optogenetic approaches are now widely used to study the function of neural populations and circuits by combining targeted expression of light-activated ...