Name: viral tracing of genetically defined neural circuitry
Uploaded: 2012-10-17T12:00:00
Description: Watch this Scientific Journal Video about Viral Tracing of Genetically Defined Neural Circuitry at JoVE.com

Vi vill tacka för Sean Whelan för hjälp med att rädda rekombinant VSV varianter och Didem Goz och Ryan Chrenek för tekniskt stöd. Detta arbete stöddes av HHMI (CLC) och # NS068012-01 (KTB).

1. Göra Virus från cDNA: Återvinning av VSV från cDNA med Vaccinia-T7-systemet 10 <ol><li> En dag före experimentet dela BsrT7 celler i 60 mm skål innehållande DMEM + 10% FBS. Seed 2E6 celler per skål. BsrT7 celler är härledda från BHK21, eller babyhamsternjurceller, celler. </li><li> Varm PBS med 1 mM magnesiumklorid och 1 mM kalcium till 37 ° C. </li><li> Skaffa en liten alikvot vTF7-3, ett vacciniavirus som uttrycker T7-polymeras, och tina till rumstemperatur. vTF7-3 är en smittsam vacciniav...

<ol>
	<li>Wickersham, I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monosynaptic+restriction+of+transsynaptic+tracing+from+single,+genetically+targeted+neurons.">Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons.</a> Neuron. 53, 639-647 (2007).</li><li>Marshel, J. H., Mori, T., Nielsen, K. J., Callaway, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+single+neuronal+networks+for+gene+expression+and+cell+labeling+in+vivo.">Targeting single neuronal networks for gene expression and cell labeling in vivo.</a> Neuron. 67, 562-574 (2010).</li><li>Wall, N. R., Wickersham, I. R., Cetin, A., De La Parra, M., Callaway, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monosynaptic+circuit+tracing+in+vivo+through+Cre-dependent+targeting+and+complementation+of+modified+rabies+virus.">Monosynaptic circuit tracing in vivo through Cre-dependent targeting and complementation of modified rabies virus.</a> Proc. Natl. Acad. Sci. U.S.A. 107, 21848-21853 (2010).</li><li>Wickersham, I. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monosynaptic+restriction+of+transsynaptic+tracing+from+single,+genetically+targeted+neurons.">Monosynaptic restriction of transsynaptic tracing from single, genetically targeted neurons.</a> Neuron. 53, 639-647 (2007).</li><li>Yonehara, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatially+asymmetric+reorganization+of+inhibition+establishes+a+motion-sensitive+circuit.">Spatially asymmetric reorganization of inhibition establishes a motion-sensitive circuit.</a> Nature. 469, 407-410 (2011).</li><li>Stepien, A. E., Tripodi, M., Arber, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monosynaptic+rabies+virus+reveals+premotor+network+organization+and+synaptic+specificity+of+cholinergic+partition+cells.">Monosynaptic rabies virus reveals premotor network organization and synaptic specificity of cholinergic partition cells.</a> Neuron. 68, 456-472 (2010).</li><li>Beier, K. T., Samson, M. E. S., Matsuda, T., Cepko, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conditional+expression+of+the+TVA+receptor+allows+clonal+analysis+of+descendents+from+Cre-expressing+progenitor+cells.">Conditional expression of the TVA receptor allows clonal analysis of descendents from Cre-expressing progenitor cells.</a> Dev. Biol. 353, 309-320 (2011).</li><li>Seidler, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Cre-loxP-based+mouse+model+for+conditional+somatic+gene+expression+and+knockdown+in+vivo+by+using+avian+retroviral+vectors.">A Cre-loxP-based mouse model for conditional somatic gene expression and knockdown in vivo by using avian retroviral vectors.</a> Proc. Natl. Acad. Sci. U.S.A. 105, 10137-10142 (2008).</li><li>Beier, K. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anterograde+or+retrograde+transsynaptic+labeling+of+CNS+neurons+with+vesicular+stomatitis+virus+vectors.">Anterograde or retrograde transsynaptic labeling of CNS neurons with vesicular stomatitis virus vectors.</a> Proc. Natl. Acad. Sci. U.S.A. 108, 15414-15419 (2011).</li><li>Whelan, S. P., Ball, L. A., Barr, J. N., Wertz, G. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+recovery+of+infectious+vesicular+stomatitis+virus+entirely+from+cDNA+clones.">Efficient recovery of infectious vesicular stomatitis virus entirely from cDNA clones.</a> Proc. Natl. Acad. Sci. U.S.A. 92, 8388-8392 (1995).</li><li>Fuerst, T. R., Niles, E. G., Studier, F. W., Moss, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Eukaryotic+Transient-Expression+System+Based+on+Recombinant+Vaccinia+Virus+That+Synthesizes+Bacteriophage+T7+RNA+Polymerase.">Eukaryotic Transient-Expression System Based on Recombinant Vaccinia Virus That Synthesizes Bacteriophage T7 RNA Polymerase.</a> PNAS. 83, 8122-8126 (1986).</li><li>Young, J. A., Bates, P., Varmus, H. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+of+a+chicken+gene+that+confers+susceptibility+to+infection+by+subgroup+A+avian+leukosis+and+sarcoma+viruses.">Isolation of a chicken gene that confers susceptibility to infection by subgroup A avian leukosis and sarcoma viruses.</a> J. Virol. 67, 1811-1816 (1993).</li><li>Madisen, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+robust+and+high-throughput+Cre+reporting+and+characterization+system+for+the+whole+mouse+brain.">A robust and high-throughput Cre reporting and characterization system for the whole mouse brain.</a> Nat Neurosci. 13, 133-140 (2010).</li><li>Franklin, K., Paxinos, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Mouse Brain in Stereotaxic Coordinates. , (1997).</li><li>van den Pol, A. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viral+strategies+for+studying+the+brain,+including+a+replication-restricted+self-amplifying+delta-G+vesicular+stomatis+virus+that+rapidly+expresses+transgenes+in+brain+and+can+generate+a+multicolor+golgi-like+expression.">Viral strategies for studying the brain, including a replication-restricted self-amplifying delta-G vesicular stomatis virus that rapidly expresses transgenes in brain and can generate a multicolor golgi-like expression.</a> J. Comp. Neurol. 516, 456-481 (2009).</li></ol>

Använda virus att studera neurala kretsar är en relativt hög genomströmning för analys anslutna neuroner. Men generera både VSV och RABV virioner är inte trivialt. Även om protokollet ovan för att rädda virus från cDNA som är det fortfarande en låg sannolikhet händelse. Nivåerna av var och en av N, P och plasmider L måste finjusteras, och många försök och replikerar behöver göras för att säkerställa viral räddning. Bildandet av ribonukleotiden partikeln innefattar en fullständig VSV-genomet RNA...

<table border="1">
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			<td>Reagent</td>
			<td>Company</td>
			<td>Catalogue number</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Tissue Culture</td>
		</tr>
		<tr>
			<td>Baby Hamster Kidney (BSRT7) cells</td>
			<td>available upon request</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>vaccinia (vTF7-3)</td>
			<td>available upon request</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>pN, pP, pl plasmids</td>
			<td>available upon request</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Calcium Chloride</td>
			<td>Sigma</td>
			<td>C1016</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Magnesium Chloride</td>
			<td>Sigma</td>
			<td>M8266</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>HEK 293T cells</td>
			<td>Open Biosystems</td>
			<td>HCL4517</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>60 mm TC-Treated Culture Dish</td>
			<td>Corning</td>
			<td>430166</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>75 cm2 Rectangular Canted Neck Cell Culture Flask with Vent Cap</td>
			<td>Corning</td>
			<td>430641</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Media : DMEM (Dulbecco's Modified Eagle Medium)</td>
			<td>Invitrogen</td>
			<td>12491-015</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>1 M HEPES pH 7.4</td>
			<td>Gibo</td>
			<td>15630-080</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>FBS: Fetal Bovine Serum</td>
			<td>Gibco</td>
			<td>10437-028</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PKS</td>
			<td>Invitrogen</td>
			<td>15140-163</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Lipofectamine 2,000 Transfection Reagent</td>
			<td>Invitrogen</td>
			<td>11668-019</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Syringe: 5 ml Luer-Lock syringe</td>
			<td>Sigma</td>
			<td>Z248010-1PAK</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Syringe Filters</td>
			<td>Nalgene</td>
			<td>190-2520</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>PEI: High Potency Linear PEI</td>
			<td>Polysciences</td>
			<td>23966</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Viral Centrifugation</td>
		</tr>
		<tr>
			<td>Corning 150 ml Tube Top Vacuum Filter System, 0.45 &mu;m Pore</td>
			<td>Corning</td>
			<td>430314</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Thinwall, Ultra-Clear, 38.5 ml, 25 x 89 mm ultracentrifuge tubes</td>
			<td>Beckman-Coulter</td>
			<td>344058</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ultracentrifuge</td>
			<td>Beckman-Coulter</td>
			<td>optima XL-80K</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>SW28 Ultracentrifuge rotor</td>
			<td>Beckman-Coulter</td>
			<td>342207</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Mouse Injection</td>
		</tr>
		<tr>
			<td>Capillary micropipets</td>
			<td>Drummond</td>
			<td>5-000-2005</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Stereotax</td>
			<td>Narishige</td>
			<td>SR-5M</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Micromanipulator</td>
			<td>Narishige</td>
			<td>SM-15</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ump injector</td>
			<td>World Precision Instruments</td>
			<td>Sys-Micro4</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Four channel microcontroller</td>
			<td>World Precision Instruments</td>
			<td>UMP3</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>M.TXB Bench Motor with C.EMX-1 Dial Control, 115 Volt</td>
			<td>Foredom</td>
			<td>M.TXB-EM</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>H.10 Handpiece, Quick Change</td>
			<td>Foredom</td>
			<td>H.10</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Step Drill, 0.5 mm</td>
			<td>Foredom</td>
			<td>A-58005P</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Microelectrode holder</td>
			<td>World Precision Instruments</td>
			<td>MEH2S</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Henry Schein</td>
			<td>995-2949</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Henry Schein</td>
			<td>4015809TV</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Henry Schein</td>
			<td>1118217</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>1 ml syringe</td>
			<td>Becton-Dickinson</td>
			<td>309628</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>30 gauge injection needle</td>
			<td>Becton-Dickinson</td>
			<td>305106</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Protective Ophthalmic Ointment</td>
			<td>Doctors Foster and Smith</td>
			<td>9N-014748</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Sigma</td>
			<td>493511</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Iodine</td>
			<td>Sigma</td>
			<td>PVP1</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Surgery and Dissection tools</td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fine Science Tools</td>
			<td>91402-12</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Standard Forceps</td>
			<td>Fine Science Tools</td>
			<td>11000-12</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Fine Forceps</td>
			<td>Fine Science Tools</td>
			<td>11255-20</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Vannas spring scissors</td>
			<td>Fine Science Tools</td>
			<td>15000-00</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Scalpel handle</td>
			<td>Fine Science Tools</td>
			<td>10003-12</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Scalpel blades</td>
			<td>Fine Science Tools</td>
			<td>10015-00</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Sutures</td>
			<td>Robbins Instruments</td>
			<td>20.SK640</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Dissection and antibody staining</td>
		</tr>
		<tr>
			<td>paraformaldehyde</td>
			<td>Sigma</td>
			<td>P6148</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline</td>
			<td>Sigma</td>
			<td>P4417</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Triton X-100</td>
			<td>Sigma</td>
			<td>T9284</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Donkey Serum</td>
			<td>Jackson Immunoresearch</td>
			<td>017-000-121</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Antibodies</td>
		</tr>
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			<td>Antibodies</td>
			<td>millipore</td>
			<td>AB144P</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Anti-gfp</td>
			<td>Abcam</td>
			<td>ab13970</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Donkey anti-chicken Dylight 488</td>
			<td>Jackson immunoresearch</td>
			<td>703-545-155</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Donkey anti-chicken Alexa Fluor 647</td>
			<td>Jackson immunoresearch</td>
			<td>705-605-147</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>DAPI</td>
			<td>Invitrogen</td>
			<td>D1306</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>&nbsp;</td>
			<td>Tissue mounting</td>
		</tr>
		<tr>
			<td>Superfrost plus microscope slides</td>
			<td>Fisher</td>
			<td>12-550-100</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Cover glass 22 x 22, 0 thickness</td>
			<td>Electron Microscopy Sciences</td>
			<td>72198-10</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Silicone elastomer</td>
			<td>Rogers Corp</td>
			<td>HT-6220</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Clear nail polish</td>
			<td>Electron Microscopy Sciences</td>
			<td>72180</td>
			<td>&nbsp;</td>
		</tr>
		<tr>
			<td>Prolong Gold antifade reagent</td>
			<td>Invitrogen</td>
			<td>P36930</td>
			<td>&nbsp;</td>
		</tr>
	</tbody>
</table>

viral tracing of genetically defined neural circuitry

Klassiska metoder för att studera neuronala kretsar är ganska låg genomströmning. Transsynaptisk virus, i synnerhet pseudorabies (PRV) och rabies virus (RABV), och på senare vesikulär stomatit virus (VSV), för att studera kretsar, blir alltmer populärt. Dessa högre genomströmning metoder använder virus som sänder mellan nervceller i antingen anterograd eller retrograd riktning. Nyligen har en modifierad RABV för monosynaptiska retrograd spårning utvecklats. (Figur 1A). I denna metod är glykoprotein (G)-genen bort från det virala genomet, och resupplied endast i riktade neuroner. Infektion specificitet uppnås genom att ersätta en chimär G, bestående av den extracellulära domänen av ASLV-Ett glykoprotein och den cytoplasmatiska domänen av RABV-G (A / RG), för den normala RABV-G 1. Denna chimära G infekterar specifikt celler som uttrycker TVA receptorn 1. Den gen som kodar TVA kan varit delivered genom olika metoder 2-8. Efter RABV-G infektion av en TVA-uttryckande neuron kan RABV överför till andra, synaptiskt anslutna nervceller i en retrograd riktning av naturen för sin egen G som var tillsammans levereras med TVA-receptorn. Denna teknik märker ett relativt stort antal ingångar (5-10%) 2 på en definierad celltyp, vilket ger ett urval av alla ingångar på en definierad förrätt celltyp. Vi har ändrats nyligen denna teknik för att använda VSV som transsynaptisk spårämne 9. VSV har flera fördelar, däribland snabbheten av genuttryck. Här har vi detaljerat ett nytt viralt system för spårning med VSV användbart för sondering microcircuitry med ökad upplösning. Även om de ursprungliga publicerade strategier för Wickersham et al. 4 och Beier et al. 9 medger märkning av alla nervceller som projekt på initialt infekterade-TVA-uttryckande-celler, här VSV var konstruerad för att överföra endast till TVA-uttryckande celler (figur 1B). Viruset först pseudotypade med RABV-G för att tillåta infektion av nervceller nedströms TVA-uttryckande neuroner. Efter infektera denna första population av celler, släpptes viruset kan bara infektera TVA-uttryckande celler. Eftersom transsynaptisk viral spridning är begränsad till TVA-uttryckande celler, närvaro eller frånvaro av anslutning från definierade celltyper kan utforskas med hög upplösning. En experimentell flödesschema av dessa experiment visas i figur 2. Här visar vi en modell krets som inriktning, selektivitet i mus näthinnan. Vi undersöker uppkopplingen av starburst amacrine celler (SAC) till näthinneganglieceller (RGC).

Watch this Scientific Journal Video about Viral Tracing of Genetically Defined Neural Circuitry at JoVE.com

Viral Tracing of Genetically Defined Neural Circuitry

En metod för att spåra synaptiskt anslutna nervceller beskrivs. Vi använder TVA specificitet en uppströms cell att söka om en cellpopulation av intresse får synaptisk input från genetiskt definierade celltyper.

Viral spåra genetiskt definierad neurala kretsar

Classical methods for studying neuronal circuits are fairly low throughput. Transsynaptic viruses, particularly the pseudorabies (PRV) and rabies virus ...

Research

JoVE Journal

Neuroscience

Physiological, Morphological and Neurochemical Characterization of Neurons Modulated by Movement

Fysiologiska, morfologiska och neurokemiska karakterisering av nervceller moduleras av Movement

The role of individual neurons and their function in neuronal circuits is fundamental to understanding the neuronal mechanisms of sensory and motor ...

Reproducible Mouse Sciatic Nerve Crush and Subsequent Assessment of Regeneration by Whole Mount Muscle Analysis

Reproducerbar Mouse ischiasnerv krossa och påföljande bedömningen av Regeneration av Whole Mount Muscle Analys

Regeneration in the peripheral nervous system (PNS) is widely studied both for its relevance to human disease and to understand the robust regenerative ...

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

Expansion of Embryonic and Adult Neural Stem Cells by In Utero Electroporation or Viral Stereotaxic Injection

Utbyggnad av embryonala och vuxna neurala stamceller genom<em&gt; I Utero</em&gt; Elektroporering eller viral stereotaxisk injektion

Somatic stem cells can divide to generate additional stem cells (expansion) or more differentiated cell types (differentiation), which is fundamental for ...

Selective Tracing of Auditory Fibers in the Avian Embryonic Vestibulocochlear Nerve

Selektiv spåra Auditiva fibrer i Avian embryonala Vestibulocochlear Nerve

The embryonic chick is a widely used model for the study of peripheral and central ganglion cell projections. In the auditory system, selective labeling ...

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

Modellering Astrocytoma Pathogenesis<em&gt; In Vitro</em&gt; Och<em&gt; In Vivo</em&gt; Använda Kortikal Astrocyter eller neurala stamceller från Villkorlig, Genmanipulerade möss

Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease ...

A Lateralized Odor Learning Model in Neonatal Rats for Dissecting Neural Circuitry Underpinning Memory Formation

En Lateralized Lukt Learning Model hos neonatala råttor för Dissecting Neural strömkrets Stödja Minnes Formation

Rat pups during a critical postnatal period (&#8804; 10 days) readily form a preference for an odor that is associated with stimuli mimicking maternal ...

Conditional Genetic Transsynaptic Tracing in the Embryonic Mouse Brain

Villkorlig Genetisk transsynaptisk Tracing i embryonal Mouse Brain

Anatomical path tracing is of pivotal importance to decipher the relationship between brain and behavior. Unraveling the formation of neural circuits ...

Laser-guided Neuronal Tracing In Brain Explants

Laserstyrda Neuronal spårning i Brain Explantat

We present a technique which combines an in vitro tracer injection protocol, which uses a series of electrical and pressure pulses to increase dye uptake ...

Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells

En-cell-RNA-Seq av definierade deluppsättningar av retinala Ganglion-celler

The discovery of cell type-specific markers can provide insight into cellular function and the origins of cellular heterogeneity. With a recent push for ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

Tidsberoende ökning av nätverkssvaret till stimulering av neuronala cellkulturer på mikroelektroder

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...