Name: an in vivo assessment of blood-brain barrier disruption in a rat model of ischemic stroke
Uploaded: 2018-03-11T14:00:00
Description: Watch this Scientific Journal Video about An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke at JoVE.com

Les auteurs sont reconnaissants à la Vice-chancellier de recherche de l’Ardabil University of Medical Sciences (Ardabil, Iran) pour le soutien financier (subvention No : 9607).

Toutes les procédures ont été effectuées conformément aux directives d’Ardabil University of Medical Sciences Research Council pour mener des études sur l’animal (numéro d’identification éthiques : IR ARUMS. Rec.1394.08). dans la présente étude visualisée, nous avons utilisé Sprague-Dawley rats adultes (300-350 g) de pâturage Institut (Téhéran, Iran).
1. anesthésie et débitmétrie
<ol><li>Induire l’anesthésie à l’isoflurane 4 % et l’entretenir à l’isofluran...

<ol>
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Jusqu’ici, diverses méthodes telles que l’autoradiographie et la détection des traceurs radioactifs24,25, immunofluorescence microscopie26,27et EB extravasation technique20, 23 ont été utilisés pour évaluer les dégâts de la barrière hémato - encéphalique. EB colorant fortement peut se lier à l’albumine sérique et est utilisé c...

Œdème cérébral vasogénique en raison de la rupture de la barrière hémato - encéphalique (BHE) reste une complication importante de l’accident vasculaire cérébral ischémique et un déterminant majeur de la survie dans les accidents vasculaires cérébraux patients1,2. La sang barrière - encéphalique (BHE), qui est formée par les cellules endothéliales capillaires du cerveau (BCECs) et composée d’éléments distincts neurovasculaire (p. ex., des jonctions serrées entre BCECs, péricytes, astroglials et les cellules neuronales3), fournit un spécialisés et dynamique de l’interface entre le système nerveux central (SNC) et la circulation sanguine périphérique4,5. Insultes tels que les lésions d’ischémie-reperfusion pourraient perturber l’intégrité fonctionnelle de la BHE et conduire à la pénétration ultérieure des leucocytes circulants dans le parenchyme du cerveau qui finalement déclencher l’inflammation cérébrale et lésions cérébrales primaires 6 , 7. modèles animaux sont nécessaires pour la détection exacte du dysfonctionnement du BBB suite à la survenue d’un accident vasculaire cérébral. Ces modèles sont d’une grande importance pour l’étude qui sous-tendent les mécanismes physiopathologiques et en introduisant de nouvelles stratégies neuroprotectrices. In vitro sur culture modèles cellulaires de la BHE hautement développées et utilisées pour l’étude moléculaire de la BBB physiopathologie8,9,10. Cependant, in vivo des modèles animaux, qui produisent des dommages ischémiques de la BHE similaire à des conditions cliniques humaines, sont également très utiles à cet égard. Détection quantitative de l’extravasation de bleu Evans (EB) est une technique bien acceptée et sensible qui a été utilisée pour évaluer l’intégrité BBB et fonction dans les maladies neurodégénératives, y compris les accidents vasculaires cérébraux ischémiques11, 12 , 13 , 14. cette méthode est rentable, réalisable, reproductible et totalement applicable dans n’importe quel laboratoire expérimental. Sa mise en œuvre ne nécessite pas d’équipements de pointe, tels que des traceurs radioactifs15 ou l’imagerie par résonance magnétique (IRM)16, qui sont indispensables pour d’autres méthodes. Dans cet article, nous démontrons exhaustive base procédés techniques d’évaluation de BBB à l’aide d’extravasation de EB dans les modèles de rat d’accident vasculaire cérébral ischémique.

<table border="0" cellpadding="0" cellspacing="0" width="829">
	<tbody>
		<tr height="22">
			<td height="22" style="height:22px;width:208px;">Isoflurane</td>
			<td style="width:203px;">Piramal</td>
			<td style="width:225px;">AWN 34041100</td>
			<td style="width:193px;">20 - 25 &#176;C</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">2,3,5-Triphenyltetrazolium chloride (TTC)</td>
			<td>Molekula</td>
			<td>31216368</td>
			<td>4 years</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Sprague&#8211;Dawley rats&#160;</td>
			<td>Pasture Institute (Tehran, Iran)</td>
			<td></td>
			<td>300-350g</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Evans Blue&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>314-13-6</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Trichloroacetic acid&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>76-03-9</td>
			<td>2 years</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Bupivacaine HCl (0.5%)</td>
			<td>Delpharm Tours</td>
			<td></td>
			<td>below&#160; 25 &#176;C</td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Bupernorphine</td>
			<td>Exir (Iran)</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Sodium Carbonate</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>497-19-8</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Sodium chloride&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>7647-14-5</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Di- Sodium hydrogen phosphate</td>
			<td>EMD Millipore&#160;</td>
			<td>231-448-7</td>
			<td></td>
		</tr>
		<tr height="23">
			<td height="23" style="height:23px;">Potassium chloride</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>&#160;7447-40-7</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:208px;">Ethanol&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>64-17-5</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">silicone(Xantopren)</td>
			<td>Heraeus</td>
			<td>EN ISO 4823</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Activator universal plus</td>
			<td>Heraeus</td>
			<td>66037445</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Micro-Dissecting forceps</td>
			<td>Stoelting</td>
			<td>52100-41</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Spring Scisors</td>
			<td>Stoelting</td>
			<td>52130-00</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Operating&#160; Scissors</td>
			<td>Roboz</td>
			<td>52140-70</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Brain matrix&#160;</td>
			<td>Stoelting</td>
			<td>51390</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Anesthesia Machine for Small Animals |</td>
			<td>&#160;Kent Scientific</td>
			<td>SS-01</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Power Lab system</td>
			<td>AD Instruments</td>
			<td>ML880</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Laser Doppler flowmeter</td>
			<td>AD Instruments</td>
			<td>ML191</td>
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			<td height="22" style="height:22px;">Heating feed back system</td>
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			<td height="22" style="height:22px;">Silk 5-0 suture thread</td>
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			<td>Ethicon</td>
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an in vivo assessment of blood-brain barrier disruption in a rat model of ischemic stroke

Accident vasculaire cérébral ischémique entraîne un œdème cérébral vasogénique et crânien primaire subséquent, qui est véhiculé par la destruction de la barrière sang - encéphalique (BHE). Rats atteints d’accident vasculaire cérébral ischémique induit ont été mis en place et utilisés comme in vivo modèles pour enquêter sur l’intégrité fonctionnelle de la BHE. Détection par spectrophotométrie de bleu Evans (EB) dans les échantillons de cerveau avec une lésion ischémique pourrait fournir une justification fiable pour la recherche et le développement de nouvelles modalités thérapeutiques. Cette méthode génère des résultats reproductibles et s’applique à n’importe quel laboratoire sans un besoin d’équipement spécial. Nous présentons ici une orientation technique et visualisée sur la détection de l’extravasation de EB après induction d’AVC ischémique chez le rat.

Il n’y avait aucune différence significative au niveau de RB dans l’hémisphère droit par rapport à l’hémisphère gauche des rats opérés (1,06 ± 0,1 µg/g et 1,1 ± 0,09 µg/g, respectivement). Comme indiqué dans les Figures 2 a-2 b, induction d’ischémie transitoire (90 min d’ischémie / reperfusion 24h) due à une différence significative au niveau de RB (10,41 ± 0,84 µg/g, p < 0,001) dans l’hémisphère gauche ischémique chez le rat, pa...

Watch this Scientific Journal Video about An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke at JoVE.com

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Une évaluation In Vivo de la perturbation de la barrière hémato - encéphalique dans un modèle de Rat d’accident vasculaire cérébral ischémique

L’objectif global de cette procédure est de fournir une technique hautement reproductible pour évaluer in vivo la rupture de la barrière hémato - encéphalique dans les modèles de rat d’accident vasculaire cérébral ischémique.

Ischemic stroke leads to vasogenic cerebral edema and subsequent primary brain injury, which is mediated through destruction of the blood-brain barrier ...

Research

JoVE Journal

Neuroscience

An In Vivo Assessment of Blood-Brain Barrier Disruption in a Rat Model of Ischemic Stroke

Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model

Generation of an Immortalized Murine Brain Microvascular Endothelial Cell Line as an In Vitro Blood Brain Barrier Model

Génération d&#39;une ligne Immortalisé microvasculaire murin des cellules endothéliales du cerveau comme un<em&gt; In Vitro</em&gt; Modèle barrière hémato-encéphalique

Epithelial and endothelial cells (EC) are building paracellular barriers which protect the tissue from the external and internal environment. The ...

Recherche

Neurosciences

Functional Neuroimaging Using Ultrasonic Blood-brain Barrier Disruption and Manganese-enhanced MRI

Utilisation de neuro-imagerie fonctionnelle par ultrasons rupture de la barrière hémato-encéphalique et le manganèse-IRM

Although mice are the dominant model system for studying the genetic and molecular underpinnings of neuroscience, functional neuroimaging in mice remains ...

Assessment of Blood-brain Barrier Permeability by Intravenous Infusion of FITC-labeled Albumin in a Mouse Model of Neurodegenerative Disease

Évaluation de la perméabilité de la barrière hémato - encéphalique par perfusion intraveineuse d’albumine marquée FITC dans un modèle murin de la maladie neurodégénérative

Disruption of blood-brain barrier (BBB) integrity is a common feature for different neurological and neurodegenerative diseases. Although the interplay ...

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

An In Vivo Blood-brain Barrier Permeability Assay in Mice Using Fluorescently Labeled Tracers

Un test In Vivo barrière hémato - encéphalique perméabilité chez la souris à l’aide de fluorescent marqué traceurs

Blood-brain barrier (BBB) is a specialized barrier that protects the brain microenvironment from toxins and pathogens in the circulation and maintains ...

A Preclinical Model to Assess Brain Recovery After Acute Stroke in Rats

Un modèle préclinique pour évaluer le rétablissement du cerveau après un AVC aigu chez les rats

The purpose of this study was to establish and validate an animal brain ischemia model in the recovery and sequela stages. A middle cerebral artery ...

A Benchtop Approach to the Location Specific Blood Brain Barrier Opening using Focused Ultrasound in a Rat Model

Une approche benchtop à l’ouverture spécifique de barrière de cerveau de sang d’emplacement utilisant l’ultrason focal dans un modèle de rat

Stereotaxic surgery is the gold standard for localized drug and gene delivery to the rodent brain. This technique has many advantages over systemic ...

Measuring Post-Stroke Cerebral Edema, Infarct Zone and Blood-Brain Barrier Breakdown in a Single Set of Rodent Brain Samples

Mesure de l’œdème cérébral post-AVC, de la zone infarctus et de la dégradation de la barrière céphalo-cérébrale dans un seul ensemble d’échantillons de cerveaux de rongeurs

One of the most common causes of morbidity and mortality worldwide is ischemic stroke. Historically, an animal model used to stimulate ischemic stroke ...

A Rat Model of EcoHIV Brain Infection

Un modèle de rat d’infection cérébrale EcoHIV

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...

Induction and Assessment of Levodopa-induced Dyskinesias in a Rat Model of Parkinson's Disease

Induction et évaluation des dyskinésies induites par la lévodopa dans un modèle rat de la maladie de Parkinson

Levodopa (L-DOPA) remains the gold-standard therapy used to treat Parkinson&#39;s disease (PD) motor symptoms. However, unwanted involuntary movements ...

Transendothelial Resistance Measurement to Assess Cell Barrier Integrity

Barrier Functional Integrity Recording on bEnd.3 Vascular Endothelial Cells via Transendothelial Electrical Resistance Detection

Applications of TEER Detection to Assess Cell Barrier Integrity (Video) | JoVE

Enregistrement de l’intégrité fonctionnelle de la barrière sur les cellules endothéliales vasculaires bEnd.3 via la détection de résistance électrique transendothéliale

The blood-brain barrier (BBB) is a dynamic physiological structure composed of microvascular endothelial cells, astrocytes, and pericytes. By coordinating ...