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

Die Autoren sind dankbar für den Vizekanzler für Forschung der Ardabil Universität der medizinischen Wissenschaften (Ardabil, Iran) für die finanzielle Unterstützung (No zu gewähren: 9607).

Alle Verfahren wurden nach den Richtlinien der Ardabil University of Medical Sciences Research Council für die Durchführung von Untersuchungen an Tieren durchgeführt (ethische ID-Nummer: IR. ARACEAE. REC.1394.08). In dieser visualisierte Studie verwendeten wir Erwachsene männlichen Sprague-Dawley Ratten (300-350 g) aus Weide Institut (Teheran, Iran) gewonnen.
1. Anästhesie und Flowmetry
<ol><li>Induzieren Sie Anästhesie mit 4 % Isofluran und pflegen Sie es mit Isofluran (1-1,5 %) in ei...

<ol>
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So weit, verschiedene Methoden wie Autoradiographie und Erkennung der radioaktiven Tracern24,25, Immunfluoreszenz-Mikroskopie26,27und EB Extravasation Technik20, 23 wurden verwendet, um die Blut - Hirn-Schranke Schaden zu bewerten. EB-Farbstoff kann sich stark an der Serum-Albumin gebunden und dient als Tracer für vaskuläre Leckagen Erkennung u...

Vasogenic Hirnödem wegen Störung der Blut - Hirn-Schranke (BBB) bleibt eine wichtige Komplikation des ischämischen Schlaganfalls und eine wichtige Determinante für die Überlebensrate in der Schlaganfall-Patienten1,2. Die Blut - Hirn-Schranke (BBB), die durch Kapillare endothelial Zellen im Gehirn (BCECs) gebildet und besteht aus verschiedenen neurovaskuläre Komponenten (z. B. enge Kreuzungen unter Perizyten, BCECs, Astroglial und neuronalen Zellen3), bietet eine spezialisierte und dynamische Schnittstelle zwischen dem zentralen Nervensystem (ZNS) und peripheren Durchblutung4,5. Beleidigungen wie Ischämie-Reperfusion Verletzungen könnte stören die funktionelle Integrität der BBB und führen nachträgliche Eindringen von zirkulierenden Leukozyten in das Gehirn Parenchym, die letztlich zerebrale Entzündung und primäre Hirnverletzungen auslösen 6 , 7. Tiermodelle für die genaue Erfassung der Dysfunktion des BBB nach Auftreten eines Schlaganfalls erforderlich sind. Solche Modelle sind von großer Bedeutung für die Untersuchung zugrunde liegenden pathophysiologische Mechanismen und Einführung von neuen neuroprotektive Strategien. In-vitro- Kultur-basierte Zellmodelle der BBB hoch entwickelt und verwendet für molekulare Studie der BBB Physiopathologie8,9,10. In Vivo Tiermodellen, die ischämische Schaden der BBB analog zu menschlichen klinischen Bedingungen zu produzieren, sind jedoch auch in dieser Hinsicht sehr lohnenswert. Quantitativen Nachweis der Extravasation von Evans blau (EB) ist eine gut angenommen und empfindliche Technik, die für die Beurteilung der BBB Integrität und Funktion bei neurodegenerativen Erkrankungen, einschließlich der ischämischen Schlaganfall11, verwendet worden ist 12 , 13 , 14. diese Methode ist kostengünstig, machbar, reproduzierbare und völlig zutreffend in einem experimentellen Labor. Ihre Umsetzung erfordert keine vorgerückte Ausrüstung, wie radioaktive Tracer15 oder Magnetresonanz-Bildgebung (MRI)16, die Voraussetzung für andere Methoden sind. In diesem Artikel zeigen wir umfassend grundlegende technische Prozesse der BBB Bewertung mit EB Extravasation in Ratte-Modelle des ischämischen Schlaganfalls.

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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>
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			<td>Molekula</td>
			<td>31216368</td>
			<td>4 years</td>
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			<td height="22" style="height:22px;">Sprague&#8211;Dawley rats&#160;</td>
			<td>Pasture Institute (Tehran, Iran)</td>
			<td></td>
			<td>300-350g</td>
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			<td>Sigma-Aldrich&#160;</td>
			<td>314-13-6</td>
			<td></td>
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			<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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			<td height="23" style="height:23px;">Bupernorphine</td>
			<td>Exir (Iran)</td>
			<td></td>
			<td></td>
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			<td height="23" style="height:23px;">Sodium Carbonate</td>
			<td>Sigma-Aldrich&#160;</td>
			<td>497-19-8</td>
			<td></td>
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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>
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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>
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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>
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			<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>
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			<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>
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			<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>
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		<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>
		<tr height="22">
			<td height="22" style="height:22px;">Heating feed back system</td>
			<td>Harvard Appratus</td>
			<td>72-7560</td>
			<td style="width:193px;"></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Vascular micro clamp</td>
			<td>FineScience Tools</td>
			<td>18055-03</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Silk 5-0 suture thread</td>
			<td>Ethicon</td>
			<td>682G</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Ethilon 4-0 suture thread&#160;</td>
			<td>Ethicon</td>
			<td>EH6740G</td>
			<td></td>
		</tr>
	</tbody>
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an in vivo assessment of blood-brain barrier disruption in a rat model of ischemic stroke

Ischämischen Schlaganfall führt zu Vasogenic zerebrales Ödem und anschließende primäre Hirnverletzung, die durch Zerstörung der Blut - Hirn-Schranke (BBB) vermittelt wird. Ratten mit induzierter ischämischen Schlaganfall wurden gegründet und als in Vivo Modellen verwendet, um die Funktionsfähigkeit der BBB zu untersuchen. Photometrische Erkennung von Evans blau (EB) in den Gehirn-Proben mit ischämischen Verletzungen konnte die Forschung und Entwicklung von neuartigen therapeutischen Modalitäten zuverlässig begründet. Diese Methode liefert reproduzierbare Ergebnisse und gilt in jedem Labor ohne spezielle Ausrüstung. Hier präsentieren wir Ihnen eine visualisierte und technische Leitlinie zur Detektion der Extravasation von EB nach Induktion der ischämische Schlaganfall bei Ratten.

Es gab keinen signifikanten Unterschied in EB Ebenen in der rechten Hemisphäre im Vergleich zu der linken Hemisphäre der Schein betrieben Ratten (1,06 ± 0,1 µg/g und 1,1 ± 0,09 µg/g, beziehungsweise). Siehe Figuren 2A-2 b, Induktion von transienten Ischämie (90 min Ischämie / Reperfusion 24 h) verursacht einen signifikanten Unterschied in EB Ebenen (10,41 ± 0,84 µg/g, p < 0,001) in der linken Hemisphäre von ischämischen Ratten, im Vergleich zu den jew...

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

Eine In-Vivo -Bewertung der Störung der Blut - Hirn-Schranke in einem Rattenmodell des ischämischen Schlaganfalls

Das übergeordnete Ziel dieses Verfahrens ist es, eine hoch reproduzierbare Technik für in Vivo Beurteilung der Blut - Hirn-Schranke Störung in Ratte-Modelle des ischämischen Schlaganfalls liefern.

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

Erzeugung eines Immortalisierte Murine Gehirn mikrovaskulären Endothelzellen Zelllinie als<em&gt; In Vitro</em&gt; Blut-Hirn-Barriere Modell

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

Forschung

Neurowissenschaften

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

Funktionelle Bildgebung mit Ultraschall-Blut-Hirnschranke Störung und Mangan-MRT

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

Beurteilung der Durchlässigkeit der Blut - Hirn-Schranke durch intravenöse Infusion von FITC-markierte Albumin in einem Mausmodell der Neurodegenerative Erkrankung

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

Eine In Vivo Blut - Hirn-Schranke Permeabilität Assay bei Mäusen mit Gewebekulturen beschriftet Tracer

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

Ein präklinisches Modell zur Beurteilung der Gehirnerholung nach akutem Schlaganfall bei Ratten

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

Ein Benchtop-Ansatz zur standortspezifischen Blut-Hirn-Barriere-Öffnung mit fokussiertem Ultraschall in einem Rattenmodell

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

Messung des Abbaus von Hirnödem nach dem Schlaganfall, Infarktzone und Blut-Hirn-Barrierenabbau in einem einzigen Satz von Nagetier-Gehirnproben

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

Ein Rattenmodell der EcoHIV-Gehirninfektion

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

Induktion und Bewertung von Levodopa-induzierten Dyskinesien in einem Rattenmodell der Parkinson-Krankheit

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

Aufzeichnung der funktionellen Barriereintegrität auf bEnd.3 vaskulären Endothelzellen mittels transendothelialer elektrischer Widerstandsdetektion

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