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

Diese Arbeit wurde von der Abteilung für auswärtige Angelegenheiten und internationalen Handel (Regierung von Kanada), University of Toronto Connaught Fund, der kanadischen Stiftung für Innovation, NSERC, der Krembil Foundation und The Cure Parkinson-Stiftung unterstützt.

1. Wohnungs-und Zubereitung von Mäusen <ol><li> Achten Sie auf eine Kolonie von Bacterial Artificial Chromosome (BAC) angetrieben transgenen Mäusen 8 (Mutant Mouse Regional Resource Center (MMRRC) FVB in einem 12:12 h Licht-Dunkel-Zyklus mit freiem Zugang zu Nahrung und Wasser. Diese Mäuse sind reine FVB-Mäusen und Es ist nicht erforderlich, diese Mäuse mit einem anderen Stamm zu überqueren, entweder zur Zucht oder für den Erfolg der 6-OHDA Läsion Verfahren zu gewährleisten. </li><li> Um eine Parki...

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Dieses Protokoll beschreibt ein Verfahren zur Erzeugung eines stabilen einseitigen 6-OHDA-Läsion Maus-Modell der Parkinson-Krankheit, die extrem reproduzierbar, mit einer hohen Erfolgsrate Läsion und eine geringe Sterblichkeit ist. Der Erfolg von 6-OHDA Läsion Operation kann leicht durch die Messung von spontanen Drehverhalten werden mit&gt; 70% ipsiversive Umdrehungen anzeigt&gt; 95% Dopaminverarmung im Striatum Läsion 27 veranschlagt. Die Quantifizierung der striatalen Dopamin-Spiegel ist die genaueste ...

<table border="1" &gt;<tbody&gt;<tr &gt;<td<strong&gt; Name des Reagenzes</strong</td&gt;<td<strong&gt; Unternehmen</strong</td&gt;<td<strong&gt; Katalog-Nummer</strong</td&gt;<td<strong&gt; Kommentare (optional)</strong</td&gt;</tr&gt;<tr &gt;<td&gt; Desipramin HCl</td&gt;<td&gt; Sigma-Aldrich, Oakville, ON, Kanada</td&gt;<td&gt; D125</td&gt;<td&gt; 25mg/kg</td&gt;</tr&gt;<tr&gt;<td&gt; Pargylin HCl</td&gt;<td&gt; Sigma-Aldrich, Oakville, ON, Kanada</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, Kanada</td&gt;<td&gt; H116</td&gt;<td&gt; 3 mg / Maus</td&gt;</tr&gt;<tr&gt;<td&gt; Stereotaktischen Rahmen</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modell 900</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Maus Ohrmuscheln</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modell 921 Zygoma Ohrenpolster</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Maus Schneidezahn bar</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modell 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Maus Narkosemaske</td&gt;<td&gt; Kopf Instruments, Tujunga, CA, USA</td&gt;<td&gt; Modell 923B</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Priming-Kit (enthält 250ml Spritze)</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 Klemmverschraubung 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 Kapillaren aus RN Klemmverschraubung Kit<
 (1/16<sup&gt; Th</sup&gt; Zoll)</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 kleinen Hub RN-Koppler</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 zu kleinen 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 Spritze Modell 25S 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 mit austauschbarer Nadel (RN) 6er Pack. . Custom 1 Zoll mit 45 &lt;° Fase</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; Scheren</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Kanada.</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 Tools, Vancouver, BC, Kanada</td&gt;<td&gt; 10003-12</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Skalpellklingen</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Kanada</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 Tools, Vancouver, BC, Kanada</td&gt;<td&gt; 11608-15</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Hämostyptika</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Kanada.</td&gt;<td&gt; 13004-14</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Isofluran</td&gt;<td&gt; Abt</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; Sterilisator</td&gt;<td&gt; Fine Science Tools, Vancouver, BC, Kanada</td&gt;<td&gt; 18000-45</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Infusionspumpe</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; Nadeln (27G)</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 305109</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Nadeln (25G)</td&gt;<td&gt; Becton Dickinson</td&gt;<td&gt; 305127</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Spritzen (1 ml)</td&gt;<td&gt; BD Spritze</td&gt;<td&gt; 309692</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Anästhesie Trolley</td&gt;<td&gt; LEI medizinische</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; 102207</td&gt;<td&gt;</td&gt;</tr&gt;<tr&gt;<td&gt; Lidocain</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 Lösung</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

Die einseitig verletzten 6-hyroxydopamine (6-OHDA)-Läsion Rattenmodell der Parkinson-Krankheit (PD) hat sich als zu einem besseren Verständnis der Mechanismen der Parkinson-Symptome von unschätzbarem Wert, da er rekapituliert die Veränderungen in den Basalganglien Schaltung und Pharmakologie in beobachtet Parkinson-Patienten 1-4. Jedoch bleiben die genauen zellulären und molekularen Veränderungen bei kortiko-striatalen Synapsen der Ausgang Wege in das Striatum, das die wichtigen Input Region der Basalganglien ist schwer zu fassen, und dies wird angenommen, dass Ort, an dem zugrunde liegenden pathologischen Auffälligkeiten Parkinson-Symptome entstehen 3 sein , 5. In PD, hat das Verständnis der Mechanismen, die Veränderungen in den Basalganglien Schaltung folgende Degeneration der nigro-striatalen Bahn wurde stark durch die Entwicklung von Bacterial Artificial Chromosome (BAC) Mäuse vorangetrieben überexprimierenden grün fluoreszierende Proteine ​​durch Förderung angetriebenRS spezifische für die beiden striatalen Bahnen Ausgang (direkter Weg: eGFP-D1; indirekten Weg: eGFP-D2 und eGFP-A2a) 8, so dass sie in Isolation untersucht werden. Zum Beispiel haben neuere Studien nahelegen, dass es pathologische Veränderungen in der synaptischen Plastizität in Parkinson-Mäusen 9,10. Diese Studien jungen Mäusen und akuten Modellen von Parkinsonismus verwendet. Es ist unklar, ob die Veränderungen bei erwachsenen Ratten, die mit stabilen 6-OHDA Läsionen beschrieben auch in diesen Modellen auftreten. Andere Gruppen haben versucht, eine stabile unilateral lädierten-6-OHDA erwachsenen Maus-Modell der PD durch Läsion des medialen Vorderhirnbündel (MFB) zu erzeugen, leider war die Mortalitätsrate in dieser Studie sehr hoch, mit nur 14% für die Operation überlebt 21 11 Tage oder länger. Neuere Studien haben intra-nigralen Läsionen sowohl mit einer niedrigen Sterblichkeitsrate generiert&gt; 80% Verlust von dopaminergen Neuronen jedoch Ausdruck von L-DOPA-induzierte Dyskinesien 11,12,13,14 war variabelIn diesen Studien. Eine andere gut etablierte Mausmodell der PD ist der MPTP-behandelten Maus 15. Während dieses Modell hat sich bewährt, bei der Beurteilung der potenziellen neuroprotektive Mittel 16 ist es weniger geeignet für das Verständnis der Mechanismen, die Symptome der PD, wie dieses Modell oft nicht gelingt, motorische Defizite zu induzieren, und zeigt eine große Variabilität in der Ausdehnung der Läsion 17, 18 . Hier haben wir eine stabile unilateralen 6-OHDA-lädierten Mausmodell der PD durch direkte Verabreichung von 6-OHDA in das MFB entwickelt, die konsequent verursacht&gt; 95% Verlust von striatalen Dopamin (gemessen durch HPLC), sowie die Herstellung der Verhaltens- Ungleichgewichte beobachtet in der Vertiefung dadurch einseitige 6-OHDA-lädierte Ratten-Modell der Parkinson. Diese neu entwickelte Mausmodell der PD wird ein wertvolles Instrument für das Verständnis der zugrunde liegenden Mechanismen Generation von Parkinson-Symptome.

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

Ein Protokoll zur Durchführung einseitige 6-OHDA Läsionen der mediale Vorderhirnbündel in Mäusen beschrieben. Dieses Verfahren hat eine niedrige Mortalitätsrate (13,3%) mit 89% der überlebenden Tiere mit&gt; 95% Verlust von striatalen Dopamin und 90,63 ± -4,02% ipsiversive Drehvorspannung nach der Seite der Läsion.

Entwicklung eines Einseitig-läsionierten 6-OHDA Mausmodell der Parkinson-Krankheit

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-Massenspektrometrie von Neuropeptiden bei Parkinsonpatienten

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

Forschung

Medizin

Skin Punch Biopsy Explant Culture for Derivation of Primary Human Fibroblasts

Haut Stanzbiopsie Explantatkultur für die Ableitung des primären humanen Fibroblasten

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

Die Verwendung von primären humanen Fibroblasten zur Überwachung Mitochondriale Phänotypen auf dem Gebiet der Parkinson-Krankheit

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

Eine einfache Methode der Maus Lung Intubation

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

Gen-Umgebung Interaktionsmodelle zur Entlarvung von Störmechanismen bei Parkinson-Krankheit

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

Controlling Parkinson-Krankheit mit Adaptive Tiefe Hirnstimulation

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

Aufbau und Charakterisierung von UTI und CAUTI in einem Mausmodell

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

A Quick Phänotypische Neurologische Scoring System zur Bewertung der Krankheitsprogression im SOD1 G93A-Mausmodell der ALS

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

Ein Neonatal Maus Spinal Cord Injury Kompression Modell

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

Entwicklung eines Alpha-Synuklein Based Rattenmodell für Parkinson-Krankheit durch stereotaktische Injektion eines rekombinanten Adeno-assoziierten viralen Vektor,

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