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

De auteurs zijn dankbaar de Vice-kanselier voor onderzoek van de Ardabil Universiteit van medische wetenschappen (Ardabil, Iran) voor de financiële ondersteuning (No verlenen: 9607).

Alle procedures werden uitgevoerd overeenkomstig de richtsnoeren van Ardabil University of Medical Sciences Research Council voor het uitvoeren van dierproeven (ethische id-nummer: IR. ARONSKELKEN. REC.1394.08). In deze gevisualiseerde studie, gebruikten we volwassen mannelijke Sprague-Dawley ratten (300-350 g) verkregen uit grasland Instituut (Teheran, Iran).
1. narcose en flowmetrie
<ol><li>Anesthesie met 4% Isofluraan induceren en onderhouden met Isofluraan (1-1,5%) in een mengsel van lac...

<ol>
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Tot nu toe diverse methoden zoals autoradiografie en opsporing van de radioactieve tracers24,25, immunofluorescentie microscopie26,27, en EB extravasation techniek20, 23 zijn gebruikt voor het evalueren van de schade van de bloed - hersenbarrière. EB kleurstof vermag sterk binden aan de serumalbumine en wordt gebruikt als een tracer voor vascula...

Vasogenic oedeem van de hersenen als gevolg van verstoring van de bloed - hersenbarrière (BBB) blijft een belangrijke complicatie van de ischemische beroerte en een belangrijke determinant van de overlevingskansen in de beroerte patiënten1,2. De bloed - hersenbarrière (BBB), die is gevormd door hersenen capillaire endotheliale cellen (BCECs) en bestaat uit verschillende neurovasculaire componenten (BV, strakke kruispunten tussen BCECs, pericytes, astroglial en neuronale cellen3), biedt een gespecialiseerde en dynamische interface tussen het centrale zenuwstelsel (CNS) en perifere bloed-circulatie4,5. Beledigingen zoals ischemie-reperfusie verwondingen kon de functionele integriteit van de BBB verstoren en leiden tot verdere penetratie van circulerende leukocyten in de hersenen parenchym die uiteindelijk leiden tot cerebrale ontsteking en primaire hersenletsel 6 , 7. dierlijke modellen nodig zijn voor de exacte detectie van de dysfunctie van BBB na optreden van een beroerte. Dergelijke modellen zijn van groot belang voor het bestuderen van de onderliggende pathofysiologische mechanismen en de invoering van nieuwe neuroprotectieve strategieën. In vitro cel cultuur gebaseerde modellen van de BBB zijn sterk ontwikkeld en gebruikt voor moleculaire studie van de BBB fysiopathologie8,9,10. In vivo diermodellen, die ischemische schade van de BBB vergelijkbaar met menselijke klinische omstandigheden produceren, zijn echter ook zeer de moeite waard in dit opzicht. Kwantitatieve detectie van de extravasation van Evans blauw (EB) is een goed aanvaard en gevoelige techniek die is gebruikt voor beoordeling van de BBB integriteit en functie in neurodegeneratieve ziekten, met inbegrip van ischemische beroerte11, 12 , 13 , 14. deze methode is kosteneffectief, haalbaar, reproduceerbaar, en volledig toepasbaar in een experimenteel laboratorium. De uitvoering ervan vereist geen geavanceerde apparatuur, zoals radioactieve tracers15 of magnetische resonantie beeldvorming (MRI)16, dat zijn voorwaarden voor andere methoden. In dit artikel tonen we uitvoerig elementaire technische processen van BBB beoordeling EB extravasation met rat modellen van ischemische beroerte.

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			<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>
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		<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>
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			<td height="23" style="height:23px;">Bupivacaine HCl (0.5%)</td>
			<td>Delpharm Tours</td>
			<td></td>
			<td>below&#160; 25 &#176;C</td>
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		<tr height="23">
			<td height="23" style="height:23px;">Bupernorphine</td>
			<td>Exir (Iran)</td>
			<td></td>
			<td></td>
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		<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>
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			<td height="23" style="height:23px;">Sodium chloride&#160;</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>7647-14-5</td>
			<td></td>
		</tr>
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			<td height="23" style="height:23px;">Di- Sodium hydrogen phosphate</td>
			<td>EMD Millipore&#160;</td>
			<td>231-448-7</td>
			<td></td>
		</tr>
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			<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>
			<td></td>
		</tr>
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an in vivo assessment of blood-brain barrier disruption in a rat model of ischemic stroke

Ischemische beroerte leidt tot vasogenic hersenoedeem en latere primaire hersenletsel, dat is bemiddeld door vernietiging van de bloed - hersenbarrière (BBB). Ratten met geïnduceerde ischemische beroerte werden opgericht en gebruikt als in vivo modellen te onderzoeken van de functionele integriteit van de BBB. Spectrofotometrische detectie van Evans blauw (EB) in de hersenen monsters met ischemische schade kon betrouwbare overleggen voor onderzoek en ontwikkeling van nieuwe therapeutische modaliteiten. Deze methode reproduceerbare resultaten oplevert, en is van toepassing in een laboratorium zonder de behoefte aan speciale apparatuur. Hier presenteren we een gevisualiseerde en technische richtsnoer op de detectie van de extravasation van EB na inductie van ischemische beroerte bij ratten.

Er was geen significant verschil in EB niveaus in de rechter hersenhelft ten opzichte van de linker hemisfeer van de sham bediende rats (1.06 ± 0,1 µg/g en 1,1 ± 0.09 µg/g, respectievelijk). Zoals in figuren 2A-2B, inductie van voorbijgaande ischemie (90 min ischemie / reperfusie 24u) veroorzaakt een significant verschil in EB niveaus (10.41 ± 0.84 µg/g, p < 0.001) in de linker hemisfeer van ischemische ratten, ten opzichte van de respectieve halfrond in de...

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

In Vivo eenbeoordeling van de verstoring van de bloed - hersen barrière in een Rat Model van ischemische beroerte

Het algemene doel van deze procedure is bedoeld als een zeer reproduceerbaar techniek voor in vivo beoordeling van de verstoring van de bloed - hersen barrière in rat modellen van ischemische beroerte.

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

Genereren van een geïmmortaliseerde Murine Brain microvasculaire endotheliale cellijn als<em&gt; In Vitro</em&gt; Blood Brain Barrier Model

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

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

Functionele neuroimaging Met behulp van ultrasone bloed-hersen barrière verstoring en mangaan-enhanced MRI

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

Beoordeling van de bloed - hersenbarrière permeabiliteit door intraveneuze infusie van FITC-geëtiketteerden albumine in een muismodel van neurodegeneratieve ziekte

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

Een In Vivo bloed - hersenbarrière permeabiliteit Assay in muizen met behulp van Fluorescently label Tracers

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

Een preklinisch model om hersen herstel na acute beroerte bij ratten te beoordelen

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

Een benchtop benadering van de locatie specifieke bloed hersenbarrière opening met behulp van gerichte echografie in een rat model

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

Het meten van post-beroerte cerebraal oedeem, infarctzone en bloed-hersenbarrière-afbraak in een enkele set knaagdierhersenmonsters

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

Een rattenmodel van EcoHIV-herseninfectie

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

Inductie en beoordeling van Levodopa-geïnduceerde dyskinesieën in een rattenmodel van de ziekte van 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

Opname van functionele integriteit van barrière op bEnd.3 vasculaire endotheelcellen via detectie van transendotheliale elektrische weerstand

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