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

우리는 재조합 VSV 변종을 구출에 도움 션 Whelan을 인정하고, 기술 지원 Didem Goz와 라이언 Chrenek 것입니다. 이 작품은 HHMI (CLC)에서 지원하고, # NS068012-01 (KTB)되었습니다.

1. cDNA에서 바이러스를 만들기 : cDNA의 VSV의 복구가 우두-T7 시스템 (10)을 사용하여 <ol><li> 실험 전에 어느 날, DMEM + 10% FBS를 포함하는 60mm 요리에 BsrT7 세포를 분리. 요리 당 종자 2E6 세포. BsrT7 세포는 BHK21, 또는 아기 햄스터 신장 세포에서 파생됩니다. </li><li> 1 ㎜의 마그네슘과 1 밀리미터 칼슘과 37 따뜻한 PBS는 ° C. </li><li> 작은 나누어지는 vTF7-3, T7의 효소를 표현 우두 바이러스 및 실내 온?...

<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>

신경 회로를 공부하고 바이러스를 사용하면 연결된 뉴런을 분석의 상대적으로 높은 처리량 방법입니다. 그러나, VSV 및 RABV의 virions 모두를 생성하는 것은 사소한되지 않습니다. cDNA에서 바이러스를 구해준 것에 대해 위에 나열된 프로토콜이 제공되고 있지만, 아직 낮은 확률 이벤트입니다. N, P 및 L의 plasmids 각각의 수준은 잘게 조정해야하고, 많은 시련과 복제는 바이러스 구조를 보장하기 위해 ?...

<table border="1">
	<tbody>
		<tr>
			<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>
		<tr>
			<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

neuronal 회로를 공부 클래식 방법은 매우 낮은 처리량 있습니다. Transsynaptic 바이러스, 회로를 공부 특히 pseudorabies (PRV)와 광견병 바이러스 (RABV), 그리고 더 최근의 기공을 갖는 구염 바이러스 (VSV)는, 점점 더 인기를 끌고있다. 이러한 높은 처리량 방법 중 하나 anterograde 또는 역행 방향으로 뉴런 사이의 전송 바이러스를 사용합니다. 최근 monosynaptic 역행 추적에 대한 수정 RABV가 개발되었습니다. (그림 1A). 이 방법에서, 당 단백질 (G) 유전자는 바이러스 게놈에서 삭제되고, 대상 뉴런에서만 resupplied. 감염 특이성은 일반 RABV-G 1 ASLV-A 당 단백질과 RABV-G (A / RG)의 세포질 도메인의 세포 도메인으로 구성된 키메라 G를, 대체에 의해 달성된다. 이 키메라 G는 특히 TVA 수용체 1을 표현 세포를 감염. 유전자 인코딩 TVA는 D되어 있습니다다양한 방법 2-8으로 elivered. TVA - 표현 뉴런의 RABV-G 감염 후, RABV는 TVA 수용체와 공동 배달 된 자신의 G의 성격에 의해 역행 방향으로 다른 synaptically 연결 뉴런에 전송할 수 있습니다. 이 기술은 정의 된 스타터 세포 유형로 입력의 모든 샘플링을 제공 정의 된 셀 타입에 입력의 상대적으로 많은 수의 (5-10 %) 2 레이블. 우리는 최근 transsynaptic 추적 9로 VSV을 사용하려면이 기술을 수정했습니다. VSV는 유전자 표현의 급속 등 여러 장점을 갖추고 있습니다. 다음은 상세 증가 해상도 microcircuitry을 탐색하는 데 유용합니다 VSV를 사용하여 새 바이러스 추적 시스템을. 동안 Wickersham 외하여 원래의 출판 전략. 4 Beier 외. 처음에 감염된로 프로젝트가 TVA--세포를 표현, 여기를 VSV는 TV 만 전송하도록 설계되었습니다있는 뉴런의 9 허가 라벨A-표현 세포 (그림 1B). 바이러스가 처음 TVA - 표현 뉴런의 뉴런 하류의 감염을 허용하는 RABV-G와 pseudotyped 있습니다. 셀의 첫 번째 인구 감염 후 바이러스는 TVA-표현 세포를 감염시킬 수 있습니다 발표했다. transsynaptic 바이러스 확산이 TVA - 표현 세포, 정의 셀 유형에서 연결의 부재의 존재로 제한되어 있기 때문에 높은 해상도로보실 수 있습니다. 이 실험의 실험 플로우 차트는 그림 2에 표시됩니다. 여기 모델 회로를 보여, 마우스 망막에 방향 선택성의. 우리는 망막 신경절 세포 (RGCs)에 항성 amacrine 세포의 연결 (주머니)를 검사합니다.

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

Viral Tracing of Genetically Defined Neural Circuitry

synaptically 연결 뉴런을 추적하는 방법이 설명되어 있습니다. 우리는 관심의 세포 인구가 유전자 정의 셀 유형에서 시냅스 입력을 받고 있는지 여부를 조사하기 위해 상류 세포의 TVA의 특이성을 사용합니다.

유 전적으로 정의 된 신경 회로의 추적 바이러스

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

운동에 의해 변조된 뉴런의 생리, 형태학의 및 Neurochemical 특성화

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

연구

신경과학

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

전체 마운트 근육 분석에 의한 재현성 마우스 좌골 신경 크러시와 중생의 후속 평가

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

의 배아와 성인 신경 줄기 세포의 확장<em&gt; 자궁에</em&gt; Electroporation 또는 바이러스 성 Stereotaxic 주입

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

조류 배아 Vestibulocochlear 신경 청각 섬유의 선택적 추적

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

모델링 성상 세포종 병인<em&gt; 체외</em&gt;와<em&gt; 생체</em&gt; 대뇌 피질의 성상이나 조건, 유전자 조작 쥐에서 신경 줄기 세포를 사용하여

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

신경 회로 토대 메모리 형성을 해부에 대한 신생아 흰쥐의 편측 냄새 학습 모델

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

배아 마우스의 뇌에 추적 조건부 유전 Transsynaptic

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

레이저 유도 신경 세포의 추적에서 뇌 외식

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

망막 신경절 세포의 정의 된 서브 세트의 단일 세포 RNA-Seq

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

미세 전극 배열에서 신경 세포 배양의 자극에 대한 네트워크 반응의 시간 의존적 증가

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