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

Questo lavoro è stato sostenuto dal Ministero degli Affari Esteri e del Commercio internazionale (Governo del Canada), University of Toronto Connaught Fondo, la Fondazione canadese per l&#39;innovazione, NSERC, la Fondazione Krembil and Trust di Parkinson Cure.

1. Housing e preparazione di topi <ol><li> Mantenere una colonia di cromosoma artificiale batterico (BAC) guidato topi transgenici 8 (Mutant mouse Regional Resource Center (MMRRC) FVB in una 12:12 h ciclo luce-buio con libero accesso a cibo e acqua. Questi topi sono topi FVB puri, e non è necessario attraversare questi topi con qualsiasi altro ceppo, sia ai fini della riproduzione, o per assicurare il successo del 6-OHDA procedura lesione. </li><li> Al fine di generare un modello parkinsoniano, topi adulti...

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Questo protocollo descrive un metodo per la generazione di uno stabile unilaterale 6-OHDA lesionato modello murino della malattia di Parkinson, che è estremamente riproducibile, con un&#39;elevata percentuale di successo lesione, e un tasso di mortalità basso. Il successo di 6-OHDA chirurgia lesione può essere facilmente stimata misurando spontaneo comportamento a rotazione con rotazioni ipsiversive&gt; 70 sulla indicativi di&gt; deplezione dopamina 95% in lesionato striato 27. Quantificazione dei livelli ...

<table border="1" &gt;<tbody&gt;<tr &gt;<td<strongNome&gt; del reagente</strong</td&gt;<td<strong&gt; Azienda</strong</td&gt;<td<strong&gt; Numero di catalogo</strong</td&gt;<td<strong&gt; Commenti (opzionale)</strong</td&gt;</tr&gt;<tr &gt;<td&gt; Desipramina 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; Pargilina 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 / mouse</td&gt;</tr&gt;<tr&gt;<td&gt; Frame stereotassico</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modello 900</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Il mouse orecchio tazze</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modello 921 coppe Zygoma</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Il mouse incisivo bar</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modello 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Il mouse anestesia maschera</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modello 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Kit di adescamento (siringa contenente 250 ml)</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 raccordo a compressione 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; Tubi PEEK dalla RN kit raccordo a compressione<
 (1/16<sup&gt; °</sup&gt; Pollici)</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 piccolo 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 ai piccoli hub RN adattatore</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; 1 ml siringa modello 25S 7001KH</td&gt;<td&gt; Hamilton Company, Reno, NV, USA</td&gt;<td&gt; 80.100</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; * 33G ago fisso (RN) confezione da 6. . Custom 1 pollice con 45 &lt;° smusso</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; Forbici</td&gt;<td&gt; Strumenti Scienza Belle, Vancouver, BC, Canada.</td&gt;<td&gt; 14084-08</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Scalpel</td&gt;<td&gt; Strumenti Scienza Arti, Vancouver, BC, Canada</td&gt;<td&gt; 10003-12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Lame di bisturi</td&gt;<td&gt; Strumenti Scienza Arti, Vancouver, BC, Canada</td&gt;<td&gt; 10035-20</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Pinza</td&gt;<td&gt; Strumenti Scienza Arti, 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; Strumenti Scienza Belle, Vancouver, BC, Canada.</td&gt;<td&gt; 13004-14</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Isoflurano</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; Sterilizzatore</td&gt;<td&gt; Strumenti Scienza Arti, Vancouver, BC, Canada</td&gt;<td&gt; 18000-45</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Infusion Pump</td&gt;<td&gt; Harvard Apparatus</td&gt;<td&gt; Dottorato di Ricerca 22/2000</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Aghi (27G)</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 305.109</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Aghi (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; Siringhe (1 ml)</td&gt;<td&gt; BD siringa</td&gt;<td&gt; 309.692</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Anestesia Carrello</td&gt;<td&gt; LEI medica</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; Lidocaina</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 soluzione</td&gt;<td&gt; CDMV, St. Hyacinthe, QC</td&gt;<td&gt; 19.955</td&gt;<td&gt;</td&gt;</tr&gt;</tbody&gt;</table

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

Il unilateralmente lesionata 6-hyroxydopamine (6-OHDA)-lesionato modello murino della malattia di Parkinson (PD) si è rivelato prezioso per avanzare la nostra comprensione dei meccanismi alla base dei sintomi parkinsoniani, poiché essa ricapitola i cambiamenti nei circuiti dei gangli della base e farmacologia osservata in pazienti parkinsoniani 1-4. Tuttavia, le modifiche precise cellulari e molecolari che si verificano a cortico-striatali sinapsi delle vie di uscita all&#39;interno del corpo striato, che è la zona di ingresso principale dei gangli basali rimanere sfuggente, e questo è ritenuto sito dove alterazioni patologiche sottostanti presentano sintomi parkinsoniani 3 , 5. Nel PD, la comprensione dei meccanismi alla base di cambiamenti nella degenerazione dei gangli basali circuito seguente del nigro-striatale è stata notevolmente migliorata con lo sviluppo del cromosoma batterico artificiale (BAC) in topi che esprimono proteine ​​fluorescenti verdi guidati da promuovere8, consentendo loro di essere valutati singolarmente:;: specifico per le due vie di uscita striatali (eGFP-D2 e eGFP-A2a via indiretta EGFP-D1 diretta via) rs. Ad esempio, studi recenti hanno suggerito che ci sono cambiamenti patologici nella plasticità sinaptica in topi parkinsoniani 9,10. Tuttavia, questi studi hanno utilizzato topi giovani e modelli acuti di parkinsonismo. Non è chiaro se le modifiche descritte in ratti adulti con stabili 6-OHDA lesioni si verificano anche in questi modelli. Altri gruppi hanno tentato di generare una stabile lesionato unilateralmente-6-OHDA modello di topo adulto del PD da parte la lesione del fascio mediale del proencefalo (MFB), purtroppo, il tasso di mortalità in questo studio è estremamente elevata, con solo il 14% sopravvive l&#39;intervento chirurgico per il 21 11 giorni o più. Studi più recenti hanno generato intra-nigral lesioni sia con un tasso di mortalità bassa perdita&gt; 80% dei neuroni dopaminergici, ma espressione di L-DOPA discinesia indotta da 11,12,13,14 era variabilein questi studi. Un altro modello di topo ben definito del PD è l&#39;MPTP-leso il mouse 15. Anche se questo modello si è dimostrato utile nella valutazione dei potenziali agenti neuroprotettivi 16, è meno adatto per la comprensione dei meccanismi alla base dei sintomi del morbo di Parkinson, in quanto questo modello spesso non riesce a indurre deficit motori, e mostra una grande variabilità nella misura di lesione 17, 18 . Qui abbiamo sviluppato una stabile unilaterale 6-OHDA lesionato modello murino di PD mediante somministrazione diretta di 6-OHDA nella MFB, che causa la perdita costante&gt; 95% della dopamina striatale (come misurato mediante HPLC), oltre a produrre il comportamento squilibri osservati nel ben caratterizzati unilaterale 6-OHDA-lesionato modello di ratto PD. Questo modello murino di recente sviluppo del PD si rivelerà un prezioso strumento per la comprensione dei meccanismi alla base della generazione dei sintomi parkinsoniani.

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

Un protocollo per l&#39;esecuzione unilaterale 6-OHDA lesioni del fascio proencefalo mediale è descritto nei topi. Questo metodo ha un basso tasso di mortalità (13,3%) con l&#39;89% degli animali sopravvissuti mostrano una perdita&gt; 95% della dopamina striatale e di 90,63 ± -4,02% pregiudizi ipsiversive rotazione verso il lato della lesione.

Sviluppo di un unilateralmente lesionata 6-OHDA modello murino della malattia di Parkinson

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

Research

JoVE Journal

Medicine

MALDI Imaging Mass Spectrometry of Neuropeptides in Parkinson's Disease

MALDI Imaging Mass Spectrometry dei neuropeptidi nella malattia di Parkinson

MALDI imaging mass spectrometry (IMS) is a powerful approach that facilitates the spatial analysis of molecular species in biological tissue samples2 ...

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Skin Punch Biopsy Explant Culture for Derivation of Primary Human Fibroblasts

Pelle Punch Biopsia Espianto Cultura per la derivazione di fibroblasti umani primari

Tissues and cell lines derived from an individual with disease are ideal sources to study disease-related cellular phenotypes. Patient-derived fibroblasts ...

The Use of Primary Human Fibroblasts for Monitoring Mitochondrial Phenotypes in the Field of Parkinson's Disease

L&#39;uso di fibroblasti umani primari per il monitoraggio fenotipi mitocondriali nel campo della malattia di Parkinson

Parkinson's disease (PD) is the second most  common movement disorder and affects 1% of people over the age of 60 1. Because ageing is  the most important ...

A Simple Method of Mouse Lung Intubation

Un metodo semplice di intubazione mouse polmone

A  simple procedure to intubate mice for pulmonary function measurements would  have several advantages in longitudinal studies with limited numbers or  ...

Gene-environment Interaction Models to Unmask Susceptibility Mechanisms in Parkinson's Disease

Modelli di interazione gene-ambiente per smascherare i meccanismi di suscettibilità nel morbo di Parkinson

Lipoxygenase (LOX) activity has been implicated in neurodegenerative disorders such as Alzheimer&#39;s disease, but its effects in Parkinson&#39;s disease ...

Controlling Parkinson's Disease With Adaptive Deep Brain Stimulation

Controllo della Malattia di Parkinson con Adaptive stimolazione cerebrale profonda

Adaptive deep brain stimulation (aDBS) has the potential to improve the treatment of Parkinson&#39;s disease by optimizing stimulation in real time ...

Establishment and Characterization of UTI and CAUTI in a Mouse Model

Istituzione e caratterizzazione di UTI e CAUTI in un modello murino

Urinary tract infections (UTI) are highly prevalent, a significant cause of morbidity and are increasingly resistant to treatment with antibiotics. ...

A Quick Phenotypic Neurological Scoring System for Evaluating Disease Progression in the SOD1-G93A Mouse Model of ALS

Un Sistema di punteggio rapida fenotipica neurologica per la valutazione di progressione della malattia nel SOD1-G93A modello murino della SLA

The SOD1-G93A transgenic mouse is the most widely used animal model of amyotrophic lateral sclerosis (ALS). At ALS TDI we developed a phenotypic screening ...

A Neonatal Mouse Spinal Cord Compression Injury Model

Un neonatale mouse Spinal Cord Injury compressione Modello

Spinal cord injury (SCI) typically causes devastating neurological deficits, particularly through damage to fibers descending from the brain to the spinal ...

Development of an Alpha-synuclein Based Rat Model for Parkinson's Disease via Stereotactic Injection of a Recombinant Adeno-associated Viral Vector

Sviluppo di un Rat modello Alfa-sinucleina per la malattia di Parkinson mediante iniezione stereotassica di un ricombinante vettore virale adeno-associato

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