Name: development of a unilaterally-lesioned 6-ohda mouse model of parkinson's disease
Uploaded: 2012-02-14T12:00:00
Description: Watch this Scientific Journal Video about Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson's Disease at JoVE.com

Dette arbeidet ble støttet av Utenriksdepartementet og International Trade (Government of Canada), University of Toronto Connaught Fund, det kanadiske Foundation for Innovation, NSERC, den Krembil Foundation og The Cure Parkinson tillit.

1. Bolig og utarbeidelse av mus <ol><li> Opprettholde en koloni av bakteriell kunstig kromosom (BAC) drevet transgene mus 8 (Mutant Mouse Regional Resource Center (MMRRC) FVB i en 12:12 h lys-mørke-syklusen med fri tilgang til mat og vann. Disse musene er rene FVB mus, og det er ikke nødvendig å krysse disse musene med andre belastninger, enten for avl, eller for å sikre suksess i 6-OHDA lesjon prosedyre. </li><li> For å generere en Parkinsonrelaterte modell, er voksne mus i alderen postnatal dag 31-42...

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Denne protokollen beskriver en metode for generering av en stabil ensidig 6-OHDA-lesioned mus modell av Parkinsons sykdom, noe som er svært reproduserbare, med en høy lesjon suksessrate, og en lav dødelighet. Suksessen av 6-OHDA lesjon kirurgi kan lett estimeres ved måling av spontan roterende adferd med&gt; 70% ipsiversive rotasjoner som tyder på&gt; 95% dopamin uttømming i lesioned striatum 27. Kvantifisering av striatale banene dopaminnivåer er den mest nøyaktige måling av omfanget av striatale ba...

<table border="1" &gt;<tbody&gt;<tr &gt;<td<strong&gt; Navn på reagensen</strong</td&gt;<td<strong&gt; Selskap</strong</td&gt;<td<strong&gt; Katalog nummer</strong</td&gt;<td<strong&gt; Kommentarer (valgfritt)</strong</td&gt;</tr&gt;<tr &gt;<td&gt; Desipramin HCl</td&gt;<td&gt; Sigma-Aldrich, Oakville, ON, Canada</td&gt;<td&gt; D125</td&gt;<td&gt; 25mg/kg</td&gt;</tr&gt;<tr&gt;<td&gt; Pargyline HCl</td&gt;<td&gt; Sigma-Aldrich, Oakville, ON, Canada</td&gt;<td&gt; P8013</td&gt;<td&gt; 5mg/kg</td&gt;</tr&gt;<tr&gt;<td&gt; 6-OHDA HBr</td&gt;<td&gt; Sigma-Aldrich, Oakville, ON, Canada</td&gt;<td&gt; H116</td&gt;<td&gt; 3 mg / mus</td&gt;</tr&gt;<tr&gt;<td&gt; Stereotaxic Ramme</td&gt;<td&gt; Kopf Instrumenter, Tujunga, CA, USA</td&gt;<td&gt; Modell 900</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Mus øret kopper</td&gt;<td&gt; Kopf Instrumenter, Tujunga, CA, USA</td&gt;<td&gt; Modell 921 Zygoma øreklokkene</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Mus fortann bar</td&gt;<td&gt; Kopf Instrumenter, Tujunga, CA, USA</td&gt;<td&gt; Modell 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Mus anestesi maske</td&gt;<td&gt; Kopf Instrumenter, Tujunga, CA, USA</td&gt;<td&gt; Modell 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Påfyllingssett (som inneholder 250ml sprøyte)</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; PRMKIT 81120</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; RN kompresjonskoblingen kit (1 mm)</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 55750-01</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; PEEK slangen fra RN kompresjonskoblingen kit<
 (1/16<sup&gt; Th</sup&gt; Tommer)</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 55751-01</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Dual liten hub RN Coupler</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 55752-01</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Luer til liten hub RN adapter</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 55753-01</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; 1ml 25s sprøyte modell 7001KH</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 80100</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; * 33G avtagbar nål (RN) pakke 6. . Custom 1 tomme med 45 &lt;° fas</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 7803-05</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Saks</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Canada.</td&gt;<td&gt; 14084-08</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Skalpell</td&gt;<td&gt; Fine Science Verktøy, Vancouver, BC, Canada</td&gt;<td&gt; 10003-12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Skalpell blad</td&gt;<td&gt; Fine Science Verktøy, Vancouver, BC, Canada</td&gt;<td&gt; 10035-20</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Forcep</td&gt;<td&gt; Fine Science Verktøy, Vancouver, BC, Canada</td&gt;<td&gt; 11608-15</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Hemostats</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Canada.</td&gt;<td&gt; 13004-14</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Isofluran</td&gt;<td&gt; Abbot</td&gt;<td&gt; 02241315</td&gt;<td&gt; 2-3%</td&gt;</tr&gt;<tr&gt;<td&gt; Suters (Vicryl 4.0)</td&gt;<td&gt; Syneture</td&gt;<td&gt; SS-683</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Sterilisatoren</td&gt;<td&gt; Fine Science Verktøy, Vancouver, BC, Canada</td&gt;<td&gt; 18000-45</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Infusjonspumpe</td&gt;<td&gt; Harvard Apparatus</td&gt;<td&gt; PhD 22/2000</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Needles (27 G)</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 305 109</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Needles (25G)</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 305 127</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Sprøyter (1ml)</td&gt;<td&gt; BD sprøyte</td&gt;<td&gt; 309 692</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Anestesi tralle</td&gt;<td&gt; LEI medisinsk</td&gt;<td&gt; M2000</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Baytril</td&gt;<td&gt; CDMV, St. Hyacinthe, QC</td&gt;<td&gt; 102 207</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Lidokain</td&gt;<td&gt; CDMV, St. Hyacinthe, QC</td&gt;<td&gt; 3914</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Betadine løsning</td&gt;<td&gt; CDMV, St. Hyacinthe, QC</td&gt;<td&gt; 19955</td&gt;<td&gt;</td&gt;</tr&gt;</tbody&gt;</table

development of a unilaterally-lesioned 6-ohda mouse model of parkinson's disease

Den ensidig lesioned 6-hyroxydopamine (6-OHDA)-lesioned rotte modell av Parkinsons sykdom (PD) har vist seg å være uvurderlig i fremme vår forståelse av mekanismene bak parkinsonsymptomer, siden det sammenfatter endringene i basalgangliene kretser og farmakologi observert i parkinsonsymptomer pasienter 1-4. Men de nøyaktige cellulære og molekylære endringer som skjer ved Cortiço-striatale banene synapser av produksjonen trasé innenfor striatum, som er den viktigste innspill regionen i basalgangliene forblir unnvikende, og dette antas å være stedet der patologiske forandringer underliggende parkinsonsymptomer oppstår 3 , 5. I PD, har forståelse for mekanismene bak endringene i basalgangliene kretser følgende degenerasjon av nigro-striatale banene veien vært sterkt frem med utviklingen av bakteriell kunstig kromosom (BAC) mus over-uttrykke grønne fluorescerende proteiner drevet av fremmers spesifikk for de to striatale banene output stier (direkte vei: eGFP-D1, indirekte vei: eGFP-D2 og eGFP-A2a) 8, slik at de kan studeres i isolasjon. For eksempel har nyere studier antydet at det er patologiske forandringer i synaptisk plastisitet i parkinsonsymptomer mus 9,10. Men disse studiene benyttes juvenile mus og akutte modeller av parkinsonisme. Det er uklart om endringene beskrevet hos voksne rotter med stabile 6-OHDA lesjoner også forekomme i disse modellene. Andre grupper har forsøkt å gi en stabil ensidig-lesioned 6-OHDA voksen mus modell av PD ved lesioning den mediale forebrain bundle (MFB), dessverre, var dødeligheten i denne studien ekstremt høy, med bare 14% overleve operasjonen for 21 dager eller lengre 11. Flere nyere studier har generert intra-nigral lesjoner med både lav dødelighet&gt; 80% tap av dopaminerge nevroner, men uttrykk for L-DOPA indusert dyskinesier 11,12,13,14 var variabeli disse studiene. Et annet godt etablert mus modell av PD er den MPTP-lesioned mus 15. Mens denne modellen har vist seg nyttig i vurderingen av mulige nervecellene agenter 16, er det mindre egnet for å forstå mekanismene bak symptomene på PD, som denne modellen ofte unnlater å indusere motor underskudd, og viser et bredt variasjon i omfanget av lesjon 17, 18 . Her har vi utviklet en stabil ensidig 6-OHDA-lesioned mus modell av PD ved direkte administrasjon av 6-OHDA i MFB, som konsekvent fører&gt; 95% tap av striatale banene dopamin (målt ved HPLC), samt produsere det atferdsmessige ubalanser observert i brønnen karakterisert ensidige 6-OHDA-lesioned rotte modell av PD. Denne nyutviklede mus modell av PD vil være et verdifullt verktøy for å forstå mekanismene bak generasjon av parkinsonsymptomer.

Watch this Scientific Journal Video about Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson's Disease at JoVE.com

Development of a Unilaterally-lesioned 6-OHDA Mouse Model of Parkinson's Disease

En protokoll for å utføre ensidige 6-OHDA lesjoner i mediale forebrain bunten i mus er beskrevet. Denne metoden har lav dødelighet (13,3%) med 89% av de overlevende dyrene viser&gt; 95% tap av striatale banene dopamin og 90,63 ± -4,02% ipsiversive roterende bias mot siden av lesjonen.

Utvikling av en ensidig-lesioned 6-OHDA mus modell av Parkinsons sykdom

The unilaterally lesioned 6-hyroxydopamine (6-OHDA)-lesioned rat model of Parkinson's disease (PD) has proved to be invaluable in advancing our ...

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In order to study the molecular pathways of Parkinson&#39;s disease (PD) and to develop novel therapeutic strategies, scientific investigators rely on ...