Name: combined in vivo anatomical and functional tracing of ventral tegmental area glutamate terminals in the hippocampus
Uploaded: 2020-09-09T12:30:00
Description: Watch this Scientific Journal Video about Combined In Vivo Anatomical and Functional Tracing of Ventral Tegmental Area Glutamate Terminals in the Hippocampus at JoVE.com

Detta arbete finansieras av CBS Bridging Grant som tilldelas OOM. OOM, PAA och AS utformade studien och utförde experimenten. AS och PAA analyserade resultaten. OOM och PAA utarbetade manuskriptet. Vi tackar Dr. Karl Disseroth (Stanford University) för att ha gjort AAV tillgängligt för vår användning.

Alla experimentella och djurhanteringsförfaranden godkändes av Institutional Animal Care and Use Committee (IACUC) vid Louisiana State University School of Veterinary Medicine.
1. Försöksdjur
<ol><li>Använd 5–6 veckor gamla möss.</li>
	<li>Hus 3-5 djur per bur under standardförhållanden på 12 h alternerande ljus och mörk cykel. Mat och vatten bör tillhandahållas ad libitum.</li>
</ol>2. Kraniotomi och djurberedning
OBS: I det h?...

<ol>
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J. <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-dependent,+anterograde+transsynaptic+viral+tracer+for+mapping+output+pathways+of+genetically+marked+neurons.">A Cre-dependent, anterograde transsynaptic viral tracer for mapping output pathways of genetically marked neurons.</a> Neuron. 72 (6), 938-950 (2011).</li><li>Li, J., Liu, T., Dong, Y., Kondoh, K., Lu, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trans-synaptic+Neural+Circuit-Tracing+with+Neurotropic+Viruses.">Trans-synaptic Neural Circuit-Tracing with Neurotropic Viruses.</a> Neuroscience bulletin. , 1-12 (2019).</li><li>Kuypers, H., Ugolini, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Viruses+as+transneuronal+tracers.">Viruses as transneuronal tracers.</a> Trends in neurosciences. 13 (2), 71-75 (1990).</li><li>Atasoy, D., Aponte, Y., Su, H. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recombinant+adeno-associated+virus+vector:+use+for+transgene+expression+and+anterograde+tract+tracing+in+the+CNS.">Recombinant adeno-associated virus vector: use for transgene expression and anterograde tract tracing in the CNS.</a> Brain Research. 793 (1-2), 169-175 (1998).</li><li>Zingg, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AAV-mediated+anterograde+transsynaptic+tagging:+mapping+corticocollicular+input-defined+neural+pathways+for+defense+behaviors.">AAV-mediated anterograde transsynaptic tagging: mapping corticocollicular input-defined neural pathways for defense behaviors.</a> Neuron. 93 (1), 33-47 (2017).</li><li>Wang, C., Wang, C., Clark, K., Sferra, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recombinant+AAV+serotype+1+transduction+efficiency+and+tropism+in+the+murine+brain.">Recombinant AAV serotype 1 transduction efficiency and tropism in the murine brain.</a> Gene therapy. 10 (17), 1528 (2003).</li><li>Ahlgrim, N. S., Manns, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optogenetic+Stimulation+of+the+Basolateral+Amygdala+Increased+Theta-Modulated+Gamma+Oscillations+in+the+Hippocampus.">Optogenetic Stimulation of the Basolateral Amygdala Increased Theta-Modulated Gamma Oscillations in the Hippocampus.</a> Frontiers in Behavioral Neuroscience. 13, 87 (2019).</li><li>Buzsaki, G., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tools+for+probing+local+circuits:+high-density+silicon+probes+combined+with+optogenetics.">Tools for probing local circuits: high-density silicon probes combined with optogenetics.</a> Neuron. 86 (1), 92-105 (2015).</li><li>Benardo, L. S., Prince, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopamine+action+on+hippocampal+pyramidal+cells.">Dopamine action on hippocampal pyramidal cells.</a> Journal of Neuroscience. 2 (4), 415-423 (1982).</li><li>Davidow, J. Y., Foerde, K., Galvan, A., Shohamy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Upside+to+Reward+Sensitivity:+The+Hippocampus+Supports+Enhanced+Reinforcement+Learning+in+Adolescence.">An Upside to Reward Sensitivity: The Hippocampus Supports Enhanced Reinforcement Learning in Adolescence.</a> Neuron. 92 (1), 93-99 (2016).</li><li>Hu, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reward+and+Aversion.">Reward and Aversion.</a> Annual Review of Neuroscience. 39, 297-324 (2016).</li><li>Kahn, I., Shohamy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intrinsic+connectivity+between+the+hippocampus,+nucleus+accumbens,+and+ventral+tegmental+area+in+humans.">Intrinsic connectivity between the hippocampus, nucleus accumbens, and ventral tegmental area in humans.</a> Hippocampus. 23 (3), 187-192 (2013).</li><li>Lisman, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relating+hippocampal+circuitry+to+function:+recall+of+memory+sequences+by+reciprocal+dentate-CA3+interactions.">Relating hippocampal circuitry to function: recall of memory sequences by reciprocal dentate-CA3 interactions.</a> Neuron. 22 (2), 233-242 (1999).</li><li>Lisman, J. E., Grace, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hippocampal-VTA+loop:+controlling+the+entry+of+information+into+long-term+memory.">The hippocampal-VTA loop: controlling the entry of information into long-term memory.</a> Neuron. 46 (5), 703-713 (2005).</li><li>Broussard, J. 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H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+rewarding+and+novel+events+in+facilitating+memory+persistence+in+a+separate+spatial+memory+task.">The role of rewarding and novel events in facilitating memory persistence in a separate spatial memory task.</a> Learning &amp; Memory. 21 (2), 61-72 (2014).</li><li>Ntamati, N. R., Luscher, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=VTA+Projection+Neurons+Releasing+GABA+and+Glutamate+in+the+Dentate+Gyrus.">VTA Projection Neurons Releasing GABA and Glutamate in the Dentate Gyrus.</a> eNeuro. 3 (4), (2016).</li><li>Yoo, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ventral+tegmental+area+glutamate+neurons+co-release+GABA+and+promote+positive+reinforcement.">Ventral tegmental area glutamate neurons co-release GABA and promote positive reinforcement.</a> Nature Communications. 7, 13697 (2016).</li><li>Funahashi, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Working+Memory+in+the+Prefrontal+Cortex.">Working Memory in the Prefrontal Cortex.</a> Brain Sciences. 7 (5), (2017).</li><li>Luo, A. H., Tahsili-Fahadan, P., Wise, R. A., Lupica, C. R., Aston-Jones, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Linking+context+with+reward:+a+functional+circuit+from+hippocampal+CA3+to+ventral+tegmental+area.">Linking context with reward: a functional circuit from hippocampal CA3 to ventral tegmental area.</a> Science. 333 (6040), 353-357 (2011).</li><li>McNamara, C. G., Dupret, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two+sources+of+dopamine+for+the+hippocampus.">Two sources of dopamine for the hippocampus.</a> Trends in Neurosciences. 40 (7), 383-384 (2017).</li><li>McNamara, C. G., Tejero-Cantero, A., Trouche, S., Campo-Urriza, N., Dupret, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dopaminergic+neurons+promote+hippocampal+reactivation+and+spatial+memory+persistence.">Dopaminergic neurons promote hippocampal reactivation and spatial memory persistence.</a> Nature Neuroscience. 17 (12), 1658-1660 (2014).</li><li>Cardin, J. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+optogenetic+stimulation+and+recording+of+neurons+in+vivo+using+cell-type-specific+expression+of+Channelrhodopsin-2.">Targeted optogenetic stimulation and recording of neurons in vivo using cell-type-specific expression of Channelrhodopsin-2.</a> Nature Protocols. 5 (2), 247-254 (2010).</li><li>Mei, Y., Zhang, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+tools+and+approaches+for+optogenetics.">Molecular tools and approaches for optogenetics.</a> Biological Psychiatry. 71 (12), 1033-1038 (2012).</li><li>Zingg, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=AAV-Mediated+Anterograde+Transsynaptic+Tagging:+Mapping+Corticocollicular+Input-Defined+Neural+Pathways+for+Defense+Behaviors.">AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways for Defense Behaviors.</a> Neuron. 93 (1), 33-47 (2017).</li><li>Zhang, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+microbial+opsin+family+of+optogenetic+tools.">The microbial opsin family of optogenetic tools.</a> Cell. 147 (7), 1446-1457 (2011).</li><li>Aravanis, A. 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Under det senaste decenniet har utformningen av AAV-konstruktioner utvecklats avsevärt. Som sådan har mer neuronspecifika promotors införlivats i en rad AAV serotyper för förbättrad transfection specificitet14. Genom att kombinera gener för fluorescensproteiner, transportörer, receptorer och jonkanaler finns nu bibliotek av AAV för avbildning, neuromodulering och synaptisk aktivitetsdetektering. I kommersiellt tillgängliga AAV-konstruktioner möjliggör en kombination av en genetiskt kod...

Under det senaste decenniet utvecklades en rad genetiska verktyg för att öka specificiteten hos neuron-typ modulering, och kartläggning av komplexa neurala nätverk1. Särskilt neurotropa virus med en inneboende förmåga att infektera och replikera i neuronala celler har distribuerats för att uttrycka eller brinna specifika proteiner i neuron subtyper. När man hyser fluorescensproteiner eller genetiskt kodade synaptiska aktivitetsindikatorer, transfecterade AAV vektorer etikett och avgränsa neurala nätverk över hjärnregioner2,3. Valet av en promotor i AAV-konstruktionen styr uttrycket av vektorn i neurontyper med en viss specificitetsnivå(promotorberoende uttryck). Men genom Cre-lox rekombination, AAV konstruktioner distribueras med större specificitet för neuron märkning4,5,6,7. Observera, fotoaktiverade mikrobiella opsinsiner och fluorescensproteiner förpackade i AAV vektorer kan uttryckas i olika neuron subtyper8, och är idealiska för bildbehandling, neuron-typ kretsspårning och fotomodulering9,10.
AAV konstruerar stereotaktiskt injiceras i en hjärnregion (eller kärna) driver uttrycket av reporterproteinet i soma, dendrite och axons terminaler. Neuralt uttryck av AAV som hyser en reportergen (eYFP) underlättar märkning av neuroncellkroppar och anatomisk spårning av projektioner till och från andrahjärnregioner 11,12,13,14. AAV-eYFP-konstruktioner som bär ljusstyrd opsin (t.ex. hChR2), kan användas som ett verktyg föravbildning 6,15 och stimuleringsbaserad fysiologisk spårning av neurala projektioner för att rikta in sig på hjärnområden in vivo16. Beroende på AAV-serotypen kan neuronmärkningens riktning vara anterograde eller bakåtsträvande11,12. Tidigare studier har visat att AAV5 reser anterogradely i nervceller12. Således ger fotostimulering av cellkroppar som uttrycker hChR2 presynaptiska effekter någon annanstans i hjärnan (målet)17.
Här användes AAV (serotyp 5) med en CaMKIIα promotor för att uttrycka eYFP (reporter) och hChR2 (opsin) i VTA glutamat nervceller och axonal projektioner. Resultaten från denna studie visar den lagerberoende fördelningen av VTA-glutamat presynaptiska terminaler i CA1, CA3 och DG hippocampal regioner. Dessutom ökade fotostimuleringen av VTA glutamatneuroner CA1 multi-unit och en enhet eldningshastigheter in vivo jämfört med baslinjevärden. Detta protokoll använder prisvärda verktyg och kommersiellt tillgänglig programvara som kan öka kvaliteten på data som erhållits från neurala kretsspårningsexperiment.

<table>
	<tbody>
		<tr>
			<td>3% Hydrogen peroxide</td>
			<td>Fisher chemical</td>
			<td>H312</td>
			<td></td>
		</tr>
		<tr>
			<td>AAV-CaMKII&#945;-ChR2-eGYP</td>
			<td>Addgene</td>
			<td>Plasmid #26969</td>
			<td></td>
		</tr>
		<tr>
			<td>BNC cable</td>
			<td>Amazon</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>BNC Splitter</td>
			<td>Amazon</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ceramic Split Mating Sleeve for &#216;1.25mm Ferrules.</td>
			<td>Thorlabs</td>
			<td>ADAL1-5</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Dremel</td>
			<td>LR 39098</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelatin coated slides</td>
			<td>Fisher scientific</td>
			<td>OBSLD01CS</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton&#39;s syringe (Neuros)</td>
			<td>WPI Inc.</td>
			<td>06H</td>
			<td></td>
		</tr>
		<tr>
			<td>Head stage adapter</td>
			<td>Neuronexus</td>
			<td>Adpt-Q4-OM32</td>
			<td></td>
		</tr>
		<tr>
			<td>High impedance silicon probe</td>
			<td>Neuronexus</td>
			<td>Q1x1-tet-5mm-121-CQ4</td>
			<td></td>
		</tr>
		<tr>
			<td>INTAN 512ch Recording Controller</td>
			<td>INTAN</td>
			<td>RHD2000</td>
			<td></td>
		</tr>
		<tr>
			<td>Iodine solution</td>
			<td>Dynarex</td>
			<td>1425</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Piramal</td>
			<td>NDC 66794-017-25</td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Spectrum</td>
			<td>K1068</td>
			<td></td>
		</tr>
		<tr>
			<td>LED Driver</td>
			<td>Thorlabs</td>
			<td>LEDD1B</td>
			<td></td>
		</tr>
		<tr>
			<td>LED light source (470 nm)-blue light</td>
			<td>Thorlabs</td>
			<td>M470F3</td>
			<td></td>
		</tr>
		<tr>
			<td>Micromanipulator</td>
			<td>Narishige</td>
			<td>M0-203</td>
			<td></td>
		</tr>
		<tr>
			<td>Optic fiber</td>
			<td>Thorlabs</td>
			<td>CFMLC14L05</td>
			<td></td>
		</tr>
		<tr>
			<td>Pan head philips screw (M0.6 X 2mm)</td>
			<td>Amazon</td>
			<td>M0.6 X 2mm</td>
			<td></td>
		</tr>
		<tr>
			<td>Pre-amplifier headstage (32 Channel)</td>
			<td>INTAN</td>
			<td>C3314</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotaxic frame</td>
			<td>Kopf</td>
			<td>1530</td>
			<td></td>
		</tr>
		<tr>
			<td>TTL pulser</td>
			<td>Prizmatix</td>
			<td>4031</td>
			<td></td>
		</tr>
		<tr>
			<td>Urethane</td>
			<td>Sigma</td>
			<td>U2500</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Alfa Aesar</td>
			<td>J61430</td>
			<td></td>
		</tr>
		<tr>
			<td>Software</td>
			<td>Company</td>
			<td>Version</td>
			<td></td>
		</tr>
		<tr>
			<td>Graphpad Prism</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Intan Recording Controller</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Neuroexplorer</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Plexon Offline Spike Sorter</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>ACSF Composition:</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>oxygenated ACSF with 95% Oxygen/5%CO2 constantly being bubbled through the ACSF (ACSF; in mM 125 NaCl, 25 NaHCO3, 3 KCl, 1.25 NaH2PO4, 1 MgCl2, 2 CaCl2 and 25 Glucose).</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

combined in vivo anatomical and functional tracing of ventral tegmental area glutamate terminals in the hippocampus

Optogenetisk modulering av neuronunderpopulationer i hjärnan har gjort det möjligt för forskare att dissekera neurala kretsar in vivo och ex vivo. Detta ger en förutsättning för att bestämma neurontypers roll inom en neural krets och deras betydelse i informationskodning i förhållande till lärande. På samma sätt kan metoden användas för att testa den fysiologiska betydelsen av två eller flera anslutna hjärnregioner hos vakna och sövda djur. Den aktuella studien visar hur VTA glutamat neuroner modulerar avfyrningshastigheten av förmodade pyramidala nervceller i CA1 (hippocampus) av sövda möss. Detta protokoll använder adeno-associerade virus (AAV)-beroende märkning av VTA glutamat nervceller för spårning av VTA presynaptic glutamat terminaler i lagren av hippocampus. Uttryck av ljusstyrd opsin (channelrhodopsin; hChR2) och fluorescensprotein (eYFP) som hyss av AAV-vektorn tillåten anterograde spårning av VTA glutamatterminaler och fotostimulering av VTA glutamat neuroncellkroppar (i VTA). Hög impedans akut kisel elektroder var placerade i CA1 för att upptäcka multi-unit och en enhet svar på VTA fotostimulering in vivo. Resultaten av denna studie visar den lagerberoende fördelningen av presynaptiska VTA glutamat terminaler i hippocampus (CA1, CA3 och DG). Dessutom ökade fototimuleringen av VTA glutamatneuroner avfyrnings- och spränghastigheten för förmodade CA1-pyramidenheter in vivo.

Spårning av anterograde
AAV uttryck kontrollerades av immunofluorescens imaging av reporter protein (eYFP) i VTA av C57BL/6 möss 21 dagar efter injektion (Figur 2). Framgångsrik anterograde märkning av presynaptic VTA glutamat projektioner i hippocampus verifierades också av eYFP detektion i lagren av DG, CA3 och CA1 (Figur 6a-d; Film 2 och 3).
VTA glutamatproj...

Watch this Scientific Journal Video about Combined In Vivo Anatomical and Functional Tracing of Ventral Tegmental Area Glutamate Terminals in the Hippocampus at JoVE.com

Combined In Vivo Anatomical and Functional Tracing of Ventral Tegmental Area Glutamate Terminals in the Hippocampus

Det aktuella protokollet visar en enkel metod för spårning av ventrala tegmental område (VTA) glutamat projektioner till hippocampus. Fotostimulering av VTA glutamat nervceller kombinerades med CA1 inspelning för att visa hur VTA glutamat terminaler modulerar CA1 förmodade pyramidal avfyrning hastighet in vivo.

Kombinerad in Vivo anatomisk och funktionell spårning av ventralt tegmentalområde glutamatterminaler i Hippocampus

Optogenetic modulation of neuron sub-populations in the brain has allowed researchers to dissect neural circuits in vivo and ex vivo. This provides a ...

Research

JoVE Journal

Neuroscience

The Analysis of Purkinje Cell Dendritic Morphology in Organotypic Slice Cultures

Analysen av Purkinje Cell Dendritic Morfologi i Organotypiska slice kulturer

Purkinje cells are an attractive model system for studying dendritic development, because they have an impressive dendritic tree which is strictly ...

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Funktionell hjärnavbildning med ultraljud Blod-hjärnbarriären störningar och mangan-förstärkt MRT

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Comprehensive Profiling of Dopamine Regulation in Substantia Nigra and Ventral Tegmental Area

Omfattande Profilering av dopamin förordning i substantia nigra och ventrala tegmentumområdet

Dopamine is a vigorously studied neurotransmitter in the CNS. Indeed, its involvement in locomotor activity and reward-related behaviour has fostered five ...

Fast Micro-iontophoresis of Glutamate and GABA: A Useful Tool to Investigate Synaptic Integration

Snabb Micro-iontophoresis av glutamat och GABA: Ett användbart verktyg för att undersöka Synaptic Integration

One of the fundamental interests in neuroscience is to understand the integration of excitatory and inhibitory inputs along the very complex structure of ...

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Ex vivo Beredningar av den Intakt vomeronasala orgel och tillbehör luktbulben

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first ...

Laser Capture Microdissection - A Demonstration of the Isolation of Individual Dopamine Neurons and the Entire Ventral Tegmental Area

Laser Capture Microdissection - En demonstration av Isolering av Individuell dopaminneuroner och hela Ventrala tegmentområdet

Laser capture microdissection (LCM) is used to isolate a concentrated population of individual cells or precise anatomical regions of tissue from tissue ...

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 ...

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Simultaneous ex vivo Functional Testing of Two Retinas by in vivo Electroretinogram System

Samtidig<em&gt; Ex vivo</em&gt; Funktionstest av två näthinnor efter<em&gt; In vivo</em&gt; Elektroretinogram System

An In vivo electroretinogram (ERG) signal is composed of several overlapping components originating from different retinal cell types, as well as noise ...

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 ...

In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin

In Vivo Functional Brain Imaging Approach Based on Bioluminescent Calcium Indicator GFP-aequorin

In Vivo Functional Brain Imaging strategi som bygger på Bioluminescent Kalcium indikator GFP-aequorin

Functional in vivo imaging has become a powerful approach to study the function and physiology of brain cells and structures of interest. Recently a new ...