Name: combined infusion and stimulation with fast-scan cyclic voltammetry (cis-fscv) to assess ventral tegmental area receptor regulation of phasic dopamine
Uploaded: 2020-04-23T14:00:00
Description: Watch this Scientific Journal Video about Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine at JoVE.c

Arbeidet ble støttet av Elizabethtown College (R.J.W, M.L., og L.M.), av et NSF Graduate Fellowship (R.J.W.) og av Yale School of Medicine (N.A.).

Alle eksperimenter ble utført i henhold til National Institutes of Health (NIH) Guide for care and use of Laboratory Animals og ble godkjent av både Elizabethtown College og Yale University Institutional Animal Care and Use Committee (IACUC). Denne protokollen er spesifikk for bedøvelse av rottepreparatet ved bruk av CIS-FSCV.
1. Presurgiske preparater
<ol><li>Forberedelse av elektrodeløsning<ol>
<li>For å lage elektrodefyllingsløsningen, lag en løsning på 4 M kaliumacetat med 140 mM...

<ol>
	<li>Grace, A. A., Bunney, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+firing+pattern+in+nigral+dopamine+neurons:+burst+firing.">The control of firing pattern in nigral dopamine neurons: burst firing.</a> Journal of Neuroscience. 4 (11), 2877-2890 (1984).</li><li>Lester, D. 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=Midbrain+acetylcholine+and+glutamate+receptors+modulate+accumbal+dopamine+release.">Midbrain acetylcholine and glutamate receptors modulate accumbal dopamine release.</a> Neuroreport. 19 (9), 991-995 (2008).</li><li>Lodge, D. J., 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+laterodorsal+tegmentum+is+essential+for+burst+firing+of+ventral+tegmental+area+dopamine+neurons.">The laterodorsal tegmentum is essential for burst firing of ventral tegmental area dopamine neurons.</a> Proceedings of the National Academy of Sciences of the United States of America. 103 (13), 5167-5172 (2006).</li><li>Li, C., 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=Mu+Opioid+Receptor+Modulation+of+Dopamine+Neurons+in+the+Periaqueductal+Gray/Dorsal+Raphe:+A+Role+in+Regulation+of+Pain.">Mu Opioid Receptor Modulation of Dopamine Neurons in the Periaqueductal Gray/Dorsal Raphe: A Role in Regulation of Pain.</a> Neuropsychopharmacology. 41 (8), 2122-2132 (2016).</li><li>Zhang, H. Y., 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=Expression+of+functional+cannabinoid+CB2+receptor+in+VTA+dopamine+neurons+in+rats.">Expression of functional cannabinoid CB2 receptor in VTA dopamine neurons in rats.</a> Addiction Biology. 22 (3), 752-765 (2017).</li><li>Wickham, R. J., 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=Advances+in+studying+phasic+dopamine+signaling+in+brain+reward+mechanisms.">Advances in studying phasic dopamine signaling in brain reward mechanisms.</a> Frontiers in Bioscience. 5, 982-999 (2013).</li><li>Wightman, R. M., 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=Monitoring+of+transmitter+metabolites+by+voltammetry+in+cerebrospinal+fluid+following+neural+pathway+stimulation.">Monitoring of transmitter metabolites by voltammetry in cerebrospinal fluid following neural pathway stimulation.</a> Nature. 262 (5564), 145-146 (1976).</li><li>Grace, A. A., Bunney, B. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+control+of+firing+pattern+in+nigral+dopamine+neurons:+single+spike+firing.">The control of firing pattern in nigral dopamine neurons: single spike firing.</a> Journal of Neuroscience. 4 (11), 2866-2876 (1984).</li><li>Mameli-Engvall, M., 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=Hierarchical+control+of+dopamine+neuron-firing+patterns+by+nicotinic+receptors.">Hierarchical control of dopamine neuron-firing patterns by nicotinic receptors.</a> Neuron. 50 (6), 911-921 (2006).</li><li>Wickham, R., 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+alpha6beta2+nicotinic+acetylcholine+receptors+modulate+phasic+dopamine+release+in+the+nucleus+accumbens+core.">Ventral tegmental area alpha6beta2 nicotinic acetylcholine receptors modulate phasic dopamine release in the nucleus accumbens core.</a> Psychopharmacology. 229 (1), 73-82 (2013).</li><li>Solecki, W., 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=Differential+role+of+ventral+tegmental+area+acetylcholine+and+N-methyl-D-aspartate+receptors+in+cocaine-seeking.">Differential role of ventral tegmental area acetylcholine and N-methyl-D-aspartate receptors in cocaine-seeking.</a> Neuropharmacology. 75, 9-18 (2013).</li><li>John, C. E., Jones, S. R., Michael, A. C., Borland, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast+Scan+Cyclic+Voltammetry+of+Dopamine+and+Serotonin+in+Mouse+Brain+Slices.">Fast Scan Cyclic Voltammetry of Dopamine and Serotonin in Mouse Brain Slices.</a> Electrochemical Methods for Neuroscience. , (2007).</li><li>Rice, M. E., Cragg, S. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nicotine+amplifies+reward-related+dopamine+signals+in+striatum.">Nicotine amplifies reward-related dopamine signals in striatum.</a> Nature Neuroscience. 7 (6), 583-584 (2004).</li><li>Espana, R. 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=Hypocretin+1/orexin+A+in+the+ventral+tegmental+area+enhances+dopamine+responses+to+cocaine+and+promotes+cocaine+self-administration.">Hypocretin 1/orexin A in the ventral tegmental area enhances dopamine responses to cocaine and promotes cocaine self-administration.</a> Psychopharmacology. 214 (2), 415-426 (2011).</li><li>Addy, N. 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=The+L-type+calcium+channel+blocker,+isradipine,+attenuates+cue-induced+cocaine-seeking+by+enhancing+dopaminergic+activity+in+the+ventral+tegmental+area+to+nucleus+accumbens+pathway.">The L-type calcium channel blocker, isradipine, attenuates cue-induced cocaine-seeking by enhancing dopaminergic activity in the ventral tegmental area to nucleus accumbens pathway.</a> Neuropsychopharmacology. 43 (12), 2361-2372 (2018).</li><li>Hermans, A., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conical+tungsten+tips+as+substrates+for+the+preparation+of+ultramicroelectrodes.">Conical tungsten tips as substrates for the preparation of ultramicroelectrodes.</a> Langmuir. 22 (25), 10348-10353 (2006).</li><li>Borland, L. M., Michael, A. C., Borland, L. M., Michael, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Introduction+to+Electrochemical+Methods+in+Neuroscience.">An Introduction to Electrochemical Methods in Neuroscience.</a> Electrochemical Methods for Neuroscience. , (2007).</li><li>Mundroff, M. L., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amperometry+and+cyclic+voltammetry+with+carbon+fiber+microelectrodes+at+single+cells.">Amperometry and cyclic voltammetry with carbon fiber microelectrodes at single cells.</a> Current Protocols in Neuroscience. 6 (6), 14 (2002).</li><li>Rodeberg, N. T., 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=Hitchhiker's+Guide+to+Voltammetry:+Acute+and+Chronic+Electrodes+for+in+vivo+Fast-Scan+Cyclic+Voltammetry.">Hitchhiker's Guide to Voltammetry: Acute and Chronic Electrodes for in vivo Fast-Scan Cyclic Voltammetry.</a> ACS Chemical Neuroscience. 8 (2), 221-234 (2017).</li><li>Sabeti, J., Gerhardt, G. A., Zahniser, N. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chloral+hydrate+and+ethanol,+but+not+urethane,+alter+the+clearance+of+exogenous+dopamine+recorded+by+chronoamperometry+in+striatum+of+unrestrained+rats.">Chloral hydrate and ethanol, but not urethane, alter the clearance of exogenous dopamine recorded by chronoamperometry in striatum of unrestrained rats.</a> Neuroscience Letters. 343 (1), 9-12 (2003).</li><li>Masuzawa, M., 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=Pentobarbital+inhibits+ketamine-induced+dopamine+release+in+the+rat+nucleus+accumbens:+a+microdialysis+study.">Pentobarbital inhibits ketamine-induced dopamine release in the rat nucleus accumbens: a microdialysis study.</a> Anesthesia &amp; Analgesia. 96 (1), 148-152 (2003).</li><li>Montague, P. R., 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=Dynamic+gain+control+of+dopamine+delivery+in+freely+moving+animals.">Dynamic gain control of dopamine delivery in freely moving animals.</a> Journal of Neuroscience. 24 (7), 1754-1759 (2004).</li><li>Keithley, R. 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=Higher+sensitivity+dopamine+measurements+with+faster-scan+cyclic+voltammetry.">Higher sensitivity dopamine measurements with faster-scan cyclic voltammetry.</a> Analytical Chemistry. 83 (9), 3563-3571 (2011).</li><li>Jackson, B. P., Dietz, S. M., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fast-scan+cyclic+voltammetry+of+5-hydroxytryptamine.">Fast-scan cyclic voltammetry of 5-hydroxytryptamine.</a> Analytical Chemistry. 67 (6), 1115-1120 (1995).</li><li>Park, J., Takmakov, P., Wightman, R. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+comparison+of+norepinephrine+and+dopamine+release+in+rat+brain+by+simultaneous+measurements+with+fast-scan+cyclic+voltammetry.">In vivo comparison of norepinephrine and dopamine release in rat brain by simultaneous measurements with fast-scan cyclic voltammetry.</a> Journal of Neurochemistry. 119 (5), 932-944 (2011).</li><li>Wenzel, J. M., 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=Phasic+Dopamine+Signals+in+the+Nucleus+Accumbens+that+Cause+Active+Avoidance+Require+Endocannabinoid+Mobilization+in+the+Midbrain.">Phasic Dopamine Signals in the Nucleus Accumbens that Cause Active Avoidance Require Endocannabinoid Mobilization in the Midbrain.</a> Current Biology. 28 (9), 1392-1404 (2018).</li><li>Spanos, M., 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=NMDA+Receptor-Dependent+Cholinergic+Modulation+of+Mesolimbic+Dopamine+Cell+Bodies:+Neurochemical+and+Behavioral+Studies.">NMDA Receptor-Dependent Cholinergic Modulation of Mesolimbic Dopamine Cell Bodies: Neurochemical and Behavioral Studies.</a> ACS Chemical Neuroscience. 10 (3), 1497-1505 (2019).</li><li>Cheer, J. 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R., 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=Optogenetic+versus+electrical+stimulation+of+dopamine+terminals+in+the+nucleus+accumbens+reveals+local+modulation+of+presynaptic+release.">Optogenetic versus electrical stimulation of dopamine terminals in the nucleus accumbens reveals local modulation of presynaptic release.</a> Journal of Neurochemistry. 134 (5), 833-844 (2015).</li><li>Sun, 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=A+Genetically+Encoded+Fluorescent+Sensor+Enables+Rapid+and+Specific+Detection+of+Dopamine+in+Flies,+Fish,+and+Mice.">A Genetically Encoded Fluorescent Sensor Enables Rapid and Specific Detection of Dopamine in Flies, Fish, and Mice.</a> Cell. 174 (2), 481-496 (2018).</li><li>Robinson, D. L., 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=Monitoring+rapid+chemical+communication+in+the+brain.">Monitoring rapid chemical communication in the brain.</a> Chemical Reviews. 108 (7), 2554-2584 (2008).</li><li>Park, J., 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=Heterogeneous+extracellular+dopamine+regulation+in+the+subregions+of+the+olfactory+tubercle.">Heterogeneous extracellular dopamine regulation in the subregions of the olfactory tubercle.</a> Journal of Neurochemistry. 142 (3), 365-377 (2017).</li><li>Ganesana, M., Venton, B. 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CIS-FSCV gir en unik mulighet til å undersøke VTA-reseptormekanismer som ligger til grunn for fasisk DA-utgivelse. Det er to kritiske trinn for å sikre riktig opptak. For det første må en stabil basislinjeopptak oppnås, med liten drift i det fremkalte DA-signalet. En viktig måte å øke sannsynligheten for å etablere et stabilt opptak er å sikre at elektroden har hatt god tid til å sykle på både 60 Hz og 10 Hz (vanligvis 15 min ved 60 Hz og 10 min ved 10 Hz). Etter hvert som karbonfiberen blir syklet, oksider...

Phasic dopamin (DA) frigjøring fra ventral tegmental området (VTA) til kjernen accumbens (NAc) spiller en viktig rolle i belønningsrelatert atferd. VTA DA nevroner bytter fra en tonic-lignende avfyring (3-8 Hz) til en burst-lignende avfyring (>14 Hz)1, som produserer fasisk DA-utgivelse i NAc. VTA uttrykker en rekke somatodendritiske reseptorer som er godt posisjonert for å kontrollere bryteren fra tonic til burst-firing2,3,4,5. Å identifisere hvilke av disse reseptorene, og deres respektive innganger, kontrollere fasisk DA-utgivelse, vil utdype vår forståelse av hvordan belønningsrelaterte kretser er organisert. Formålet med metodikken som er beskrevet her, kombinert infusjon og stimulering med hurtigskanning av syklisk voltammetri (CIS-FSCV), er å raskt og robust vurdere funksjonaliteten til VTA-reseptorer i å drive fasisk DA-frigjøring.
Begrepet kombinert infusjon og stimulering (CIS) refererer til farmakologisk manipulerende reseptorer på en gruppe nevroner (her VTA) og stimulerer disse nevronene til å studere reseptorens funksjon. I den bedøvede rotten stimulerer vi VTA elektrisk for å fremkalle et stort fasisk DA-signal (1-2 μM) i NAc-kjernen, målt ved hurtigskanning av syklisk voltammetri (FSCV). Infusjoner av farmakologiske legemidler (dvs. reseptoragonister/antagonister) på stimuleringsstedet kan brukes til å måle funksjonen til VTA-reseptorer ved å observere den påfølgende endringen i fremkalt fasisk DA-frigjøring. FSCV er en elektrokjemisk tilnærming som har både høy romlig (50-100 μm) og temporal (10 Hz) oppløsning, og er godt egnet til å måle belønningsrelaterte, fasiske DA-hendelser6,7. Denne oppløsningen er finere enn andre in vivo nevrokjemiske målinger, for eksempel mikrodialyse. Sammen er CIS-FSCV derfor velegnet til å vurdere VTA-reseptorregulering av fasisk dopaminfrigjøring.
En vanlig måte å undersøke VTA-reseptorfunksjonen på er ved å bruke en kombinasjon av elektrofysiologiske tilnærminger som adresserer hvordan disse reseptorene endrer avfyringshastigheten til nevroner1,8. Disse studiene er svært verdifulle for å forstå hvilke reseptorer som er involvert i å drive DA-avfyring ved aktivering. Imidlertid kan disse studiene bare foreslå hva som kan skje nedstrøms ved axonterminalen (dvs. frigjøring av en nevrotransmitter). CIS-FSCV bygger på disse elektrofysiologiske studiene ved å svare på hvordan utgangen av VTA burst-firing, phasic DA release, reguleres av reseptorer som ligger på VTA dendritter og cellelegemer. Dermed er CIS-FSCV godt egnet til å bygge på disse elektrofysiologistudiene. Som et eksempel kan nikotinreseptoraktivering indusere burst-firing i VTA9, og CIS-FSCV i bedøvelsesrotten ble brukt til å vise at nikotinisk acetylkolinreseptor (nAChR) aktivering i VTA også kontrollerer fasisk DA-utgivelse i NAc10,11.
Mekanistisk undersøkelse av fasisk DA-regulering studeres også ofte ved hjelp av skiver preparater sammen med bad påføring av legemidler. Disse studiene fokuserer ofte på presynaptisk regulering av phasic DA-frigjøring fra dopaminterminaler, da cellelegemene ofte fjernes fra skive12. Disse preparatene er verdifulle for å studere presynaptiske reseptoreffekter på dopaminterminaler, mens CIS-FSCV er bedre egnet til å studere somatodendritiske reseptoreffekter på dopaminnevroner, samt presynaptiske innganger til VTA. Dette skillet er viktig, fordi somatodendritisk reseptoraktivering i VTA kan ha en annen effekt enn NAc presynaptisk reseptoraktivering. Faktisk kan blokkering av dopaminerge presynaptiske nAChRs i NAc heve fasic dopamin frigjøring under burst-firing13, mens det motsatte er sant på VTA somatodendritc nAChRs10,11.
CIS-FSCV er en ideell tilnærming for å studere VTA-reseptorers evne til å regulere fasisk DA-frigjøring. Det er viktig at denne tilnærmingen utføres i en intakt rotte, enten bedøvet eller fri bevegelig. Denne tilnærmingen er egnet for akutte studier, for å studere reseptorfunksjon i baseline tilstand10,14 samt langsiktige studier som kan vurdere funksjonelle endringer i en reseptor etter legemiddeleksponering eller atferdsmanipulering11,15.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Electrode Filling Solution/Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micropipette</td>
			<td>World Precision Instruments</td>
			<td>MF286-5 (28 gauge)</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Acetate</td>
			<td>Sigma</td>
			<td>236497-100G</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Chloride</td>
			<td>Sigma</td>
			<td>P3911-25G</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Carbon fiber</td>
			<td>Thornel</td>
			<td>T650</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode puller</td>
			<td>Narishige International</td>
			<td>PE-22</td>
			<td>Note: horizontal pullers can be used as well</td>
		</tr>
		<tr>
			<td>Glass capillary</td>
			<td>A-M systems</td>
			<td>626000</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulated wires for electrodes</td>
			<td>Weico Wire and Cable Incorporated</td>
			<td>UL 1423</td>
			<td>Length; 10 cm; diameter,0.4mm; must get custom made; insulated material should cover 5 cm of the wire</td>
		</tr>
		<tr>
			<td>Light Microscope (for viewing and cutting electrode)</td>
			<td>Fischer Scientific</td>
			<td>M3700</td>
			<td></td>
		</tr>
		<tr>
			<td>Pin</td>
			<td>Phoenix Enterprises</td>
			<td>HWS1646</td>
			<td>To be soldered onto the insuled electrode wire and reference electrode; connects to headstage</td>
		</tr>
		<tr>
			<td>Putty</td>
			<td>Alcolin</td>
			<td><a target="_blank" href="https://www.dickblick.com/items/23922-1003/">23922-1003</a></td>
			<td>Used to place electrode on while cutting the carbon fiber</td>
		</tr>
		<tr>
			<td>Scalpal Blade</td>
			<td>World Precision Instruments</td>
			<td>500239</td>
			<td>For cutting carbon fiber to the apprpriate length</td>
		</tr>
		<tr>
			<td>Silver Wire</td>
			<td>Sigma</td>
			<td>327026-4G</td>
			<td></td>
		</tr>
		<tr>
			<td>FSCV Hardware/Software</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Faraday Cage</td>
			<td>U-Line</td>
			<td>H-3618 (36&#34; x 24&#34; x 42&#34;)</td>
			<td></td>
		</tr>
		<tr>
			<td>Potentiostat</td>
			<td>Univ. of N. Carolina, Electronics Facility</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stimulating electrode</td>
			<td>PlasticsOne</td>
			<td>MS303/2-A/SPC</td>
			<td>when ordering, request a 22 mm cut below pedestal</td>
		</tr>
		<tr>
			<td>TarHeel HDCV Software</td>
			<td>University of North Carolina-Chapel Hill</td>
			<td>-</td>
			<td><a href="https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/" target="_blank">https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/</a> for ordering information</td>
		</tr>
		<tr>
			<td>UEI breakout box</td>
			<td>Univ. of N. Carolina, Electronics Facility</td>
			<td></td>
			<td><a href="https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/" target="_blank">https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/</a> for ordering information</td>
		</tr>
		<tr>
			<td>UEI power supply</td>
			<td>Univ. of N. Carolina, Electronics Facility</td>
			<td></td>
			<td><a href="https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/" target="_blank">https://chem.unc.edu/critcl-main/criticl-electronics/criticl-electronics-hardware/</a> for ordering information</td>
		</tr>
		<tr>
			<td>Stimulator Hardware</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Neurolog stimulus isolator</td>
			<td>Digitimer Ltd.</td>
			<td>DS4</td>
			<td>Neurolog 800A</td>
		</tr>
		<tr>
			<td>Infusion/Stimulation Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Infusion Pump</td>
			<td>New Era Syringe Pump</td>
			<td>NE-300</td>
			<td></td>
		</tr>
		<tr>
			<td>Internal Cannula</td>
			<td>PlasticsOne</td>
			<td>C315I/SPC INTERNAL 33GA</td>
			<td></td>
		</tr>
		<tr>
			<td>Microliter Syringe</td>
			<td>Hamilton</td>
			<td>80308</td>
			<td></td>
		</tr>
		<tr>
			<td>Tubing</td>
			<td>PlasticsOne</td>
			<td>C313CT/ PKG TUBING 023 X 050 PE50</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Cannula Holder</td>
			<td>Kopf Instruments</td>
			<td>1776 P-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton Tip Applicators</td>
			<td>Vitality Medical</td>
			<td>806</td>
			<td></td>
		</tr>
		<tr>
			<td>Electrode Holder</td>
			<td>Kopf Instruments</td>
			<td>1770</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating Pad</td>
			<td>Kent Scientific</td>
			<td>RT-0501</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Vitality Medical</td>
			<td>29906-004</td>
			<td></td>
		</tr>
		<tr>
			<td>Screws</td>
			<td>Stoelting</td>
			<td>Bone Anchor Screws/Pkg.of 100</td>
			<td>1.59 mm O.D., 3.2 mm long</td>
		</tr>
		<tr>
			<td>Silver wire reference with AgCl</td>
			<td>InVivo Metric</td>
			<td>E255A</td>
			<td></td>
		</tr>
		<tr>
			<td>Square Gauze</td>
			<td>Vitality Medical</td>
			<td>441408</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotax</td>
			<td>Kopf Instruments</td>
			<td>Model 902 (Dual Arm Bar)</td>
			<td></td>
		</tr>
		<tr>
			<td>Histological Supplies</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Formulin</td>
			<td>Sigma</td>
			<td>1004960700</td>
			<td></td>
		</tr>
		<tr>
			<td>Power supply</td>
			<td>BK Precision</td>
			<td>9110</td>
			<td></td>
		</tr>
		<tr>
			<td>Sucrose</td>
			<td>Sigma</td>
			<td>80497</td>
			<td></td>
		</tr>
		<tr>
			<td>Tungsten microelectrode</td>
			<td>MicroProbes</td>
			<td>WE30030.5A3</td>
			<td></td>
		</tr>
		<tr>
			<td>Drugs for infusions</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>((2R)-amino-5-phosphonovaleric acid</td>
			<td>Sigma Aldrich</td>
			<td>A5282</td>
			<td></td>
		</tr>
		<tr>
			<td>N-methyl-D-aspartate</td>
			<td>Sigma Aldrich</td>
			<td>M3262</td>
			<td></td>
		</tr>
		<tr>
			<td>Mecamylamine hydrochloride (M9020-5mg)</td>
			<td>Sigma Aldrich</td>
			<td>M9020</td>
			<td></td>
		</tr>
		<tr>
			<td>Scopolamine hydrobromide (S0929-1g)</td>
			<td>Sigma Aldrich</td>
			<td>S0929</td>
			<td></td>
		</tr>
	</tbody>
</table>

combined infusion and stimulation with fast-scan cyclic voltammetry (cis-fscv) to assess ventral tegmental area receptor regulation of phasic dopamine

Phasic dopamin (DA) frigjøring fra ventral tegmental området (VTA) til kjernen accumbens spiller en sentral rolle i belønning behandling og forsterkning læring. Å forstå hvordan de forskjellige nevroninngangene i VTA-kontrollens fasiske DA-utgivelse kan gi et bedre bilde av kretsene som styrer belønningsbehandling og forsterkningslæring. Her beskriver vi en metode som kombinerer intra-VTA cannula infusjoner av farmakologiske agonister og antagonister med stimulering-fremkalt fasisk DA-frigjøring (kombinert infusjon og stimulering, eller CIS) målt ved in vivo hurtigskanning syklisk voltammetri (FSCV). Ved hjelp av CIS-FSCV hos bedøvede rotter kan en fasisk DA-respons fremkalles ved å stimulere VTA elektrisk med en bipolar elektrode utstyrt med en kanyle mens du registrerer i kjernen av kjernen. Farmakologiske agonister eller antagonister kan infunderes direkte på stimuleringsstedet for å undersøke spesifikke VTA-reseptorers roller i å drive fasisk DA-frigjøring. En stor fordel med CIS-FSCV er at VTA-reseptorfunksjonen kan studeres in vivo, basert på in vitro-studier.

CIS-FSCV ble brukt til å studere funksjonen til VTA N-metyl-D-aspartatreseptorer (NMDAR), nikotiniske acetylkolinreseptorer (nAChRs) og muskariniske acetylkolinreseptorer (mAChRs) i drivende fasisk DA-frigjøring i NAc-kjernen. Figur 2 viser representative data for en negativ kontroll, infusjon av 0,9 % saltvann, før (baseline) og 9 min postinfusjon (saltvann). Figur 2 viser et fargeplott med potensial på y-aksen, tid på x-ak...

Watch this Scientific Journal Video about Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine at JoVE.c

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry CIS-FSCV to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

Målet med denne protokollen er å direkte manipulere ventrale tegmentale områdereseptorer for å studere deres bidrag til subsecond dopaminfrigjøring.

Kombinert infusjon og stimulering med hurtigskanning av syklisk voltammetri (CIS-FSCV) for å vurdere ventral tegmental områdereseptorregulering av fasisk dopamin

Phasic dopamine (DA) release from the ventral tegmental area (VTA) to the nucleus accumbens plays a pivotal role in reward processing and reinforcement ...

Research

JoVE Journal

Neuroscience

Combined Infusion and Stimulation with Fast-Scan Cyclic Voltammetry (CIS-FSCV) to Assess Ventral Tegmental Area Receptor Regulation of Phasic Dopamine

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network

The default mode network is a group of brain regions that are active when an individual is not focused on the outside world and the brain is at "wakeful ...

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry

Presynaptiske Dopamin Dynamics i striatale banene Brain Slices med Fast-scan Syklisk Voltammetry

Extensive research has focused on the neurotransmitter dopamine because of its importance in the mechanism of action of drugs of abuse (e.g. cocaine and ...

Voltage Biasing, Cyclic Voltammetry, &amp; Electrical Impedance Spectroscopy for Neural Interfaces

Voltage Biasing, Cyclic Voltammetry, & Electrical Impedance Spectroscopy for Neural Interfaces

Spenning administratorinnstilling, Syklisk Voltammetry, og elektrisk impedans spektroskopi for Neural Interfaces

Electrical impedance spectroscopy (EIS) and cyclic voltammetry (CV) measure properties of the electrode-tissue interface without additional invasive ...

Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry

After its release into the synaptic cleft, dopamine exerts its biological properties via its pre- and post-synaptic targets1. The dopamine signal is ...

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

Omfattende profilering av Dopamin forordning i substantia nigra og Ventral tegmentale området

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

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

Laser Capture mikrodisseksjon - En demonstrasjon av Isolering av Individuell dopaminneuroner og hele Ventral tegmentale området

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

The Preparation of Oblique Spinal Cord Slices for Ventral Root Stimulation

Utarbeidelse av Oblique Spinal Cord Slices for Ventralt Root Stimulering

Electrophysiological recordings from spinal cord slices have proven to be a valuable technique to investigate a wide range of questions, from cellular to ...

Modeling Fast-scan Cyclic Voltammetry Data from Electrically Stimulated Dopamine Neurotransmission Data Using QNsim1.0

Modellering Fast-scan-syklusvoltammetridata fra elektrostimulert data for dopamin-neurotransmisjon ved bruk av QNsim1.0

Central dopaminergic (DAergic) pathways have an important role in a wide range of functions, such as attention, motivation, and movement. Dopamine (DA) is ...

Effects of Transcranial Alternating Current Stimulation on the Primary Motor Cortex by Online Combined Approach with Transcranial Magnetic Stimulation

Effekter av Transkraniell vekselstrøm stimulering på primære motorisk Cortex av Online kombinert tilnærming med Transkraniell magnetisk stimulering

Transcranial Alternating Current Stimulation (tACS) is a neuromodulatory technique able to act through sinusoidal electrical waveforms in a specific ...