Præsynaptiske dopamin Dynamics i striatale Brain Skiver med Fast-scan Cycliske voltammetri
Summary
Brug af Fast-scan cyklisk voltammetri at måle elektrisk fremkaldte præsynaptiske dopamin dynamik i striatale hjernen skiver.
Abstract
Omfattende forskning har fokuseret på neurotransmitteren dopamin grund af dens betydning i virkningsmekanismen af misbrugsstoffer (fx kokain og amfetamin), den rolle, den spiller i psykiatriske lidelser (fx skizofreni og Attention Deficit Hyperactivity Disorder), og dets engagement i degenerative lidelser som Parkinsons og Huntingtons sygdom. Under normale fysiologiske forhold, er dopamin kendt for at regulere motorisk aktivitet, kognition, læring, følelsesmæssig affekt og neuroendokrine hormon sekretion. En af de største tætheder af dopamin neuroner i striatum, som kan opdeles i to adskilte neuroanatomiske regioner kaldet nucleus accumbens og caudatus, putamen. Formålet er at illustrere en generel protokol for skive hurtigt scan cyklisk voltammetri (FSCV) inden musen striatum. FSCV er et veldefineret elektrokemisk teknik giver mulighed for at måle dopamin frigivelse og optagelse i realtid i discrete hjernen regioner. Kulfiber mikroelektroder (diameter ~ 7 μm) anvendes i FSCV at opdage dopamin oxidation. Den analytiske fordel ved at bruge FSCV at opdage dopamin er dens forbedret tidsmæssige opløsning på 100 millisekunder og rumlig opløsning på mindre end ti mikrometer, der giver supplerende oplysninger til in vivo mikrodialyse.
Protocol
Discussion
Den protokol, der præsenteres her viser hvordan man udarbejder og bruge musen koronale hjernen skiver for FSCV eksperimenter. Selv om denne metode er specifik for at opnå og måle dopamin dynamik, andre neurotransmittere som adenosin, er hydrogenperoxid, noradrenalin og serotonin blevet overvåget i vivo eller in vitro med FSCV 3, 8 – 11. FSCV kan bruges til at overvåge nogle af disse andre neurochemicals ved simple ændringer i bølgeform anvendes på arbejde elektrode 3, 11. …
Disclosures
The authors have nothing to disclose.
Acknowledgements
Finansiering fra National Institute for alkoholmisbrug og alkoholisme (NIAAA; AA-016967 og AA016967-01S1, TAM), start Wayne State University op fonde, og Wayne State University Research Grant Program. Indholdet er alene forfatternes ansvar og repræsenterer ikke det officielle synspunkter NIAAA eller National Institutes of Health.
Materials
| Name | Company | CAS number |
Reagent |
||
Potassium Chloride |
Fisher | 7447407 |
Sodium chloride |
EMD Chemicals | 7647145 |
Magnesium chloride |
Fisher |
7791186 |
Calcium chloride |
Fisher |
10035048 |
Sodium bicarbonate |
EMD Chemicals |
144558 |
Sodium phosphate,Dibasic |
EMD Chemicals |
7558794 |
D-glucose |
Fisher |
50997 |
Ascorbic acid |
Fisher |
50817 |
Sucrose |
Fisher |
57501 |
|
|
|
Materials |
Vendor |
Catalogue number |
Carbon fiber |
Goodfellow Oakdale, PA | |
Glass capillary |
A-M Systems Carlsborg WA. | 602000 |
Silver wire |
A-M Systems Carlsborg WA. | 787000 |
Tungsten stimulating electrode |
Plastics One, Roanoke, VA | |
Platinum wire |
||
Lead wire |
Squires Electronics, Cornelius, OR | |
Loctite 404 instant adhesive |
Hankal Corp. Rocky Hill CT. |
|
Razor blade |
World precision Instruments Inc. FL. | |
BD Spinal needle |
BD Medical systems, Franlin Lake, NJ |
REF 405234 |
Surgical Blade |
Feather Safety razor Co. LTD. Japan | |
|
||
Software |
Vendor |
Catalogue number |
TH software |
ESA Inc.,Chelmsford, MA | |
Instrument |
Vendor |
Catalogue number |
Submersion recording chamber |
Custom Scientific, Denver, CO | |
Neorolog stimulus isolator |
Digitmeter, Hertfordshire, England | |
Automatic temperature controller |
Warner Instrument Corporation | |
Microscope (SZX7) |
Olympus | |
Microscope |
Fisher | |
Vibratome 3000 sectioning system |
St. Louis , MO. | |
Perfusion pump |
Watson Marlow Limited, Falmouth Cornwall – England | H110708 |
Micropipette puller |
Narishige, Tokyo, Japan | |
ChemClamp potentiostat |
Dagan Corporations, Minneapolis, MN. |
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