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

Dette arbeidet ble støttet av livsforsikringsselskap &quot;Novalis-Taitbout&quot;, Ecole des Neurosciences de Paris (ENP), den Agence Nationale de la Recherche &quot;ANR-09-NEUR-004&quot; i rammen av &quot;ERA-NET neuron&quot; av FP7 program av Europakommisjonen, &quot;Foundation pour la Recherche Medicale&quot;, den Letten Foundation og Pasteur Foundation.

1. Stereotaxic injeksjoner <ol><li> Viruset som brukes i disse forsøkene er en replikering-mangelfull lentiviral vektor basert på HIV-viruset å uttrykke Channelrhodopsine2-YFP (gule fluorescerende protein) fusion konstruere drevet av en synapsin promoter 6. Plasmidet ble sjenerøst gitt av K. Deisseroth &#39;s forskergruppe ( <a href="http://www.stanford.edu/group/dlab/optogenetics/index.html">http://www.stanford.edu/group/dlab/optogenetics/index.html</a> ). Lentiviruses kan fås fra kommersielle kilder ...

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Optogenetic verktøy nylig har økt vår kontroll over selektiv nevronal befolkningen, inkludert voksen født nevroner i luktsystemet. Her beskriver vi hvordan du utfører en stereotaxic injeksjon av en lentiviral vektor uttrykker ChR2-YFP i en bestemt populasjon av voksen-fødte nevroner. Ved å nøye overvåke perioden etter injeksjon, er vi i stand til å analysere og manipulere aktiviteten til en kohort av voksen-fødte nevroner med en relativ homogen alder. På grunn av deres lave diffu...

<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> Glass Kapillærer (3,5 &quot;) </td><td> Drummond Scientific </td><td> 3-000-203-G / X </td></tr><tr><td> Nanoject II Microinjector </td><td> Drummond Scientific </td><td> 3-000-204 </td></tr><tr><td> Høy hastighet kirurgisk drill </td><td> Fin Science Verktøy </td><td> 18000-17 </td></tr><tr><td> Micro Bor Steel grader 0.7mm tip diameter </td><td> Fin Science Verktøy </td><td> 19008-07 </td></tr><tr><td> LED CMS blå 4,6 W </td><td> OSRAM </td><td> LBW5SN </td></tr><tr><td> Kvinne elektrisk kontakt (2,54 mm) </td><td> Fischer Elektronik </td><td> BL5.36Z </td></tr><tr><td> Nåværende controller for LED (max: 1A) </td><td> A1W Electronik </td><td> HKO-KL50-1000 </td></tr><tr><td> Pulsgeneratorens </td><td> AMPI </td><td> Master-8 </td></tr><tr><td> Optisk strømmåler </td><td> Thor Labs </td><td> PM100 </td></tr><tr><td> Cyanoakrylat lim </td><td> Loctite </td><td> 454 </td></tr><tr><td> Polycarboxylate sement (første lag sement) </td><td> Septodont </td><td> Selfast (# 0068Q) </td></tr><tr><td> Methacrylate sement (andre lag sement) </td><td> GC InternationalCorp. </td><td> Unifast Trad - Liquid (# 339292) Unifast Trad - Powder (# 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 interneurons kontinuerlig regenereres i luktelappen hos voksne gnagere 1-3. I denne prosessen kalles voksen neurogenesis, nevrale stamceller i veggene i den laterale ventrikkelen gi opphav til neuroblasts som vandrer for flere millimeter langs rostral vandrende stream (RMS) for å nå og innlemme i luktelappen. Å studere de ulike trinnene og virkningen av voksen-født neuron integrering i preexisting olfactory kretser, er det nødvendig å selektivt etiketten og manipulere aktiviteten av denne bestemte befolkning av nerveceller. Den siste tidens utvikling av optogenetic teknologier gir muligheten til å bruke lys til å nettopp aktivere denne spesifikke kohort av nerveceller uten å påvirke omkringliggende nerveceller 4,5. Her presenterer vi en rekke prosedyrer for å virally uttrykke Channelrhodopsin2 (ChR2)-YFP i en timelig begrenset kohort av neuroblasts i RMS før de når luktelappen og bli voksen-født neurons. I tillegg viser vi hvordan implantatet og kalibrere en miniatyr LED for kronisk in vivo stimulering av ChR2-uttrykke nevroner.

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

Voksen-fødte nevroner i luktelappen kan optogenetically styres ved hjelp Channelrhodopsin2-uttrykke lentiviral injeksjon i rostral vandrende bekken og kronisk photostimulation med en implantert miniatyr LED.

Selektiv Viral Transduksjon of Adult-fødte olfactory Neurons for Chronic In vivo Optogenetic Stimulering

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

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 ...

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

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

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 Fotolyse av Caged forbindelser i Cilia av olfactory sensoriske nevroner

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

Fiberoptisk Implantasjon for kronisk Optogenetic Stimulering av hjernevev

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

Optogenetic Stimulering av Escape Behavior i<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

En metode for High Fidelity optogenetic kontroll av enkeltPyramideFormet Neuroner<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 ...

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Whole Mount Immunolabeling of Olfactory Receptor Neurons in the Drosophila Antenna

Hele Mount immunolabeling av olfactory Receptor nevroner i<em&gt; Drosophila</em&gt; Antenne

Odorant molecules bind to their target receptors in a precise and coordinated manner. Each receptor recognizes a specific signal and relays this ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo optogenetic Stimulering av gnagere sentralnervesystemet

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

Intracerebroventrikulær Viral Injeksjon av Neonatal Mouse Brain for Vedvarende og Utbredt Neuronal Transduksjon

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

Optogenetic Stimulering av hørselsnerven

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