Name: presynaptic dopamine dynamics in striatal brain slices with fast-scan cyclic voltammetry
Uploaded: 2012-01-12T13:00:00
Description: Watch this Scientific Journal Video about Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry at JoVE.com

Finansiering levert av National Institute on Alcohol Abuse og alkoholisme (Niaa; AA-016967 og AA016967-01S1, TAM), Wayne State University starte opp midler, og Wayne State University Research Grant Program. Innholdet er utelukkende ansvaret til forfattere og representerer ikke offisielle utsikt over Niaa eller National Institutes of Health.

1. Eksperimentell Essentials Elektrode Fabrication <ul type="circle"><li> Det er mange karbonfiber microelectrodes fabrikasjon metoder siden de fleste er laget på huset. Vanligvis hva dikterer elektroden fabrikasjon detaljer er elektrokjemiske teknikken som er brukt til elektroden (f.eks amperometry vs FSCV). For FSCV kan microelectrodes gjøres i huset ved hjelp av følgende tre-trinns prosedyre. For en mer fullstendig beskrivelse av karbon fiber elektrode fabrikasjon, ser at det nylig Jove...

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Protokollen som presenteres her demonstrerer hvordan å forberede og bruke musen koronale hjernen skiver for FSCV eksperimenter. Selv om denne metoden er spesifikk for innhenting og måling dopamin dynamikk, andre nevrotransmittere som adenosin, har hydrogen peroxide, noradrenalin og serotonin vært overvåket in vivo eller in vitro med 3 FSCV, 8-11. FSCV kan brukes til å overvåke noen av disse andre nevrokjemiske med enkle modifikasjoner av bølgeform brukes til å arbeide elektrode ...

<table border="1"><tbody><tr><td> Navn </td><td> Firma </td><td> CAS-nummer </td></tr><tr><td width="274"> Reagens </td><td></td><td></td></tr><tr><td width="274"> Kaliumklorid </td><td> Fisher </td><td width="142" valign="top"> 7447407 </td></tr><tr><td width="274"> Natriumklorid </td><td> EMD Kjemikalier </td><td width="142" valign="top"> 7647145 </td></tr><tr><td width="274"> Magnesiumklorid </td><td width="194" valign="top"> Fisher </td><td width="142" valign="top"> 7791186 </td></tr><tr><td width="274"> Kalsiumklorid </td><td width="194" valign="top"> Fisher </td><td width="142" valign="top"> 10035048 </td></tr><tr><td width="274"> Natriumbikarbonat </td><td width="194" valign="top"> EMD Kjemikalier </td><td width="142" valign="top"> 144558 </td></tr><tr><td width="274"> Natriumfosfat, Dibasic </td><td width="194" valign="top"> EMD Chemicaler </td><td width="142" valign="top"> 7558794 </td></tr><tr><td width="274"> D-glukose </td><td width="194" valign="top"> Fisher </td><td width="142" valign="top"> 50997 </td></tr><tr><td width="274"> Askorbinsyre </td><td width="194" valign="top"> Fisher </td><td width="142" valign="top"> 50817 </td></tr><tr><td width="274"> Sukrose </td><td width="194" valign="top"> Fisher </td><td> 57501 </td></tr><tr><td width="274">&nbsp;</td><td width="194" valign="top">&nbsp;</td><td>&nbsp;</td></tr><tr><td width="274"> Materialer </td><td width="194" valign="top"> Vendor </td><td> Katalognummer </td></tr><tr><td width="274"> Carbon fiber </td><td> Goodfellow Oakdale, PA </td><td></td></tr><tr><td width="274"> Glass kapillær </td><td> AM Systems Carlsborg WA. </td><td> 602000 </td></tr><tr><td width="274"> Silver ledning </td><td> A-M Systems Carlsborg WA. </td><td> 787000 </td></tr><tr><td width="274"> Tungsten stimulerende elektrode </td><td> Plast One, Roanoke, VA </td><td></td></tr><tr><td width="274"> Platinatråd </td><td></td><td></td></tr><tr><td width="274"> Ledning </td><td> Squires Electronics, Cornelius, OR </td><td></td></tr><tr><td width="274"> Loctite 404 instant lim </td><td> Hankal Corp Rocky Hill CT. </td><td></td></tr><tr><td width="274"> Barberblad </td><td> Verden presisjon Instruments Inc. FL. </td><td></td></tr><tr><td width="274"> BD Spinal nål </td><td> BD Medical Systems, Franlin Lake, NJ </td><td> NR 405234 </td></tr><tr><td width="274"> Kirurgisk Blade </td><td> Feather barberhøvel Co LTD. Japan </td><td></td></tr><tr><td width="274"></td><td></td><td></td></tr><tr><td width="274"> Software </td><td width="194" valign = &quot;top&quot;&gt; Vendor </td><td> Katalognummer </td></tr><tr><td width="274"> TH programvare </td><td> ESA Inc., Chelmsford, MA </td><td></td></tr><tr><td width="274"> Instrument </td><td width="194" valign="top"> Vendor </td><td> Katalognummer </td></tr><tr><td width="274"> Neddykking innspilling kammer </td><td> Custom Scientific, Denver, CO </td><td></td></tr><tr><td width="274"> Neorolog stimulus isolator </td><td> Digitmeter, Hertfordshire, England </td><td></td></tr><tr><td width="274"> Automatisk temperatur controller </td><td> Warner Instrument Corporation </td><td></td></tr><tr><td width="274"> Mikroskop (SZX7) </td><td> Olympus </td><td></td></tr><tr><td width="274"> Microscope </td><td> Fisher </td><td></td></tr><tr><td width="274"> Vibratome 3000 seksjonering system </td><td> St. Louer, MO. </td><td></td></tr><tr><td width="274"> Perfusjon pumpe </td><td> Watson Marlow Limited, Falmouth Cornwall - England </td><td> H110708 </td></tr><tr><td width="274"> Mikropipette puller </td><td> Narishige, Tokyo, Japan </td><td></td></tr><tr><td width="274"> ChemClamp potentiostat </td><td> Dagan Corporations, Minneapolis, MN. </td><td></td></tr></tbody></table>

presynaptic dopamine dynamics in striatal brain slices with fast-scan cyclic voltammetry

Omfattende forskning har fokusert på signalstoffet dopamin grunn av dens betydning i virkningsmekanismen til narkotika av misbruk (f.eks kokain og amfetamin), hvilken rolle den spiller i psykiatriske sykdommer (for eksempel schizofreni og Attention Deficit Hyperactivity Disorder), og sitt engasjement i degenerative lidelser som Parkinsons og Huntingtons sykdom. Under normale fysiologiske forhold, er dopamin kjent for å regulere locomotor aktivitet, kognisjon, læring, følelsesmessig påvirke, og nevroendokrin hormon. En av de største tetthetene av dopamin nevroner er innenfor striatum, som kan deles i to atskilte nevroanatomi områder kalt nucleus accumbens og caudatus-putamen. Målet er å illustrere en generell protokoll for skive hurtig-scan syklisk voltammetry (FSCV) innenfor musen striatum. FSCV er en veldefinert elektrokjemiske teknikken gir mulighet til å måle dopamin frigjøring og opptak i sanntid i discrete hjernen regioner. Carbon fiber microelectrodes (diameter på ~ 7 mikrometer) brukes i FSCV å oppdage dopamin oksidasjon. Den analytiske Fordelen med å bruke FSCV å oppdage dopamin er dens forbedret temporal oppløsning på 100 millisekunder og romlig oppløsning på mindre enn ti mikrometer, gir komplementær informasjon til in vivo microdialysis.

Watch this Scientific Journal Video about Presynaptic Dopamine Dynamics in Striatal Brain Slices with Fast-scan Cyclic Voltammetry at JoVE.com

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

Ved hjelp av rask-scan syklisk voltammetry å måle elektrisk fremkalt presynaptiske dopamin dynamikk i striatale banene hjernen skiver.

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 ...

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