Name: intravital microscopy and thrombus induction in the earlobe of a hairless mouse
Uploaded: 2017-04-02T14:00:00
Description: Watch this Scientific Journal Video about Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse at JoVE.com

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Alle in vivo-experimenten (7221.3-1-006 / 15) werden uitgevoerd in overeenstemming met de Duitse wetgeving inzake de bescherming van dieren en de NIH Gids voor de Zorg en gebruik van proefdieren (Institute of Laboratory Animal Resources, National Research Council). 1. Algemene houden van de dieren <ol><li> Voer de experimenten met mannelijke SKH1-Hr uur muizen van 4 tot 6 weken. Dieren worden gebruikt met een gewicht tussen 20 en 25 g. </li><li> Bewaar de dieren in een ...

<ol>
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Er zijn een aantal cruciale stappen voor de succesvolle trombus inductie in de oorlel van SKH1-Hr hr muizen. Voor het oplossen van problemen, worden de respectievelijke stappen van het protocol aangegeven tussen haakjes. Onderzoek omstandigheden zijn ideaal bij jonge dieren op de leeftijd van 4-6 weken en met een lage verhoorning van de opperhuid. Bij oudere dieren, de kwaliteit van visualisatie van de vaten slechter en meer vergelijkbaar vanwege de hogere afstand tussen het huido...

Het doel van dit artikel is de techniek van intravitale microscopie toegepast op de oorschelp van de haarloze muis voor de directe observatie en analyse van microcirculatie, trombusvorming en trombus evolutie beschrijven. Met een incidentie van 1 op 1000, veneuze trombose nog steeds een veel voorkomende oorzaak van morbiditeit. Hoewel diagnostiek, preventiestrategieën en therapieën zijn in de afgelopen jaren zijn ontwikkeld, een derde van de veneuze trombose manifesteert zich als een longembolie 1. Arteriële trombose speelt een cruciale rol in hart- en vaatziekten, die de meest voorkomende doodsoorzaak in geïndustrialiseerde landen zijn. Arteriële trombose op basis van de breuk van atherosclerotische plaques is betrokken bij hartaanvallen, mesenteriale infarcten, en beroerte. Elke operatie bloot subendotheliale structuren bloedcomponenten, verandert de dynamiek van de bloedstroom, en immobiliseert de patiënt. In endoprosthetische operatie aan de onderste ledematen, orgel transplantation en flap chirurgie trombose zijn frequente oorzaken van complicaties. Microvasculaire trombose in het bijzonder veroorzaakt vaak onherstelbare schade, als gevolg van het gebrek aan klinische symptomen. Ook microvasculaire trombose speelt een cruciale regel in verschillende ziekten, waaronder trombotische trombocytopenische purpura, sepsis, diffuse intravasale stolling, antifosfolipidensyndroom en veneuze insufficiëntie, onder anderen. Verschillende nieuwe geneesmiddelen voor de behandeling en preventie van trombose werden ontwikkeld in de afgelopen jaren, maar antiplatelet drugs en antistollingsmiddelen hebben nog steeds bijwerkingen, gebrek antagonisten, en zijn voorzien van lange duur effecten. Deze tekortkomingen leiden tot problemen in dringende medische zorg. Aldus is meer onderzoek nodig om de complexe processen die optreden tijdens trombose, die nauwelijks kan worden gesimuleerd in vitro bloot te leggen. De haarloze SKH1-Hr hr muis werd ontdekt 1926 in een dierentuin in Londen.Vanwege een gendefect op chromosoom 14, het dier verliest zijn vacht na postnatale dag 10. Dit geeft een goed gevasculariseerde auricula toegankelijk intravitale microscopie van de vaten. De gemiddelde dikte van het oor is 300 urn. Het bestaat uit twee lagen van de dermis, die worden gescheiden door kraakbeen. Aan de convexe dorsale zijde van het kraakbeen, 3 vaatbundels voert de oorlel. Apicale vasculaire bogen en basale shunts sluit de drie bundels. De venules hebben diameters tussen 200 urn (basaal) en 10 urn (apicale). Fijnmazig haarvaten rond de lege haarzakjes 2. De anatomie van de haarloze SKH1-Hr hr muis maakt de oorschelp een krachtige en kosteneffectieve model voor trombose onderzoek.

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			<td height="19" style="height:19px;width:216px;">SKH-1/hr mice</td>
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			<td style="width:161px;">Aesculap</td>
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			<td>can be purchased from other vendors&#160;</td>
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			<td>can be purchased from other vendors&#160;</td>
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		<tr height="19">
			<td height="19" style="height:19px;width:216px;">microscissors</td>
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			<td style="width:231px;">OC496R</td>
			<td>can be purchased from other vendors&#160;</td>
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		<tr height="19">
			<td height="19" style="height:19px;width:216px;">scalpel 21</td>
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			<td>can be purchased from other vendors&#160;</td>
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			<td height="19" style="height:19px;width:216px;">Prolene 7-0</td>
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			<td style="width:231px;">XNEH7470</td>
			<td>can be purchased from other vendors&#160;</td>
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			<td height="19" style="height:19px;width:216px;">Prolene 6-0</td>
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			<td height="19" style="height:19px;width:216px;">electrocautery</td>
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</table>

intravital microscopy and thrombus induction in the earlobe of a hairless mouse

Trombotische complicaties van vasculaire ziekten zijn een belangrijke oorzaak van morbiditeit en mortaliteit in de geïndustrialiseerde landen. Vanwege de complexe interacties tussen cellulaire en niet-cellulaire bloedcomponenten bij thrombusvorming, kan betrouwbare studies van de fysiologie en pathofysiologie van trombose alleen worden uitgevoerd in vivo. Daarom worden in dit artikel een oor model in haarloze muizen en richt zich op de in vivo analyse van de microcirculatie, trombusvorming en trombus evolutie. Via intravitale fluorescentiemicroscopie en intraveneuze (iv) toepassing van de respectieve fluorescerende kleurstoffen, kan een herhaalde analyse van de microcirculatie in de oorschelp gemakkelijk worden uitgevoerd, zonder de noodzaak van chirurgische voorbereiding. Bovendien kan dit model worden aangepast voor in vivo studies van verschillende zaken, zoals wondgenezing, reperfusieletsel, of angiogenese. Kortom, het oor van haarloze muizen is een ideaal model voor de in vivo onderzoek microcirculatie in fysiologische of pathofysiologische aandoeningen en de evaluatie van de reactie op andere systemische of topische behandelingen.

Effecten van Cannabinoid Treatment thrombogenese Na injectie van 0,05 ml FITC-dextran, fototoxisch trombus inductie leidt tot een endotheliale laesie en de vorming van een pariëtale plaatjesprop (figuren 2 en 3). In deze studie thrombus-inductie na ip injectie van cannabinoïden (5 mg / kg lichaamsgewicht) of vehikel tot een trombotische occlusie vat alle venules (figuur 4). Bij...

Watch this Scientific Journal Video about Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse at JoVE.com

Intravital Microscopy and Thrombus Induction in the Earlobe of a Hairless Mouse

Het oor model van de kale SKH1-Hr uur muis maakt intravitale fluorescentiemicroscopie microcirculatie en fototoxische trombus inductie zonder voorafgaande chirurgische preparaten in de onderzochte microvasculaire bed. Daarom is het oor van de haarloze muis is een uitstekend in vivo model voor de complexe interacties te bestuderen tijdens microvasculaire trombusvorming, trombus evolutie, en trombolyse.

Opklaren en Trombus inductie in de oorlel van een Hairless Mouse

Thrombotic complications of vascular diseases are one leading cause of morbidity and mortality in industrial nations. Due to the complex interactions ...

Research

JoVE Journal

Medicine

Technical Aspects of the Mouse Aortocaval Fistula

Technische aspecten van de muis Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

Pharmacologic Induction of Epidermal Melanin and Protection Against Sunburn in a Humanized Mouse Model

Farmacologische Inductie van epidermale melanine en bescherming tegen zonnebrand in een gehumaniseerd muismodel

Fairness of skin, UV sensitivity and skin cancer  risk all correlate with the physiologic function of the melanocortin 1  receptor, a Gs-coupled signaling ...

An Orthotopic Glioblastoma Mouse Model Maintaining Brain Parenchymal Physical Constraints and Suitable for Intravital Two-photon Microscopy

Een orthotope Glioblastoma Muis Model Handhaving Brain Parenchymale fysieke beperkingen en geschikt voor intravitale twee-foton microscopie

Glioblastoma multiforme (GBM) is the most aggressive form of brain tumors with no curative treatments available to date.
Murine models of this pathology ...

Intravital Video Microscopy Measurements of Retinal Blood Flow in Mice

Intravitale Video Microscopie Metingen van retinale bloedstroom in Muizen

Alterations in retinal blood flow can contribute to, or be a consequence of, ocular disease and visual dysfunction. Therefore, quantitation of altered ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Inductie van versnelde atherosclerose bij muizen: het "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Combined Intravital Microscopy and Contrast-enhanced Ultrasonography of the Mouse Hindlimb to Study Insulin-induced Vasodilation and Muscle Perfusion

Gecombineerde Intravital microscopie en Contrast-enhanced Ultrasonografie van het stuk van de muis om te bestuderen van de insuline-geïnduceerde vasodilatatie en spier perfusie

It has been demonstrated that insulin's vascular actions contribute to regulation of insulin sensitivity. Insulin's effects on muscle perfusion regulate ...

Inducing Ischemia-reperfusion Injury in the Mouse Ear Skin for Intravital Multiphoton Imaging of Immune Responses

Induceren van ischemie-reperfusie schade in de Mouse Ear Skin voor intravitale Multiphoton Imaging van immuunresponsen

Ischemia-reperfusion injury (IRI) occurs when there is transient hypoxia due to the obstruction of blood flow (ischemia) followed by a subsequent ...

Intravital Microscopy of Monocyte Homing and Tumor-Related Angiogenesis in a Murine Model of Peripheral Arterial Disease

Intravital microscopie van monocyt Homing en Tumor-gerelateerde angiogenese in een lymfkliertest Model van perifere arteriële ziekte

The therapeutic goal for peripheral arterial disease and ischemic heart disease is to increase blood flow to ischemic areas caused by hemodynamic ...

Induction and Micro-CT Imaging of Cerebral Cavernous Malformations in Mouse Model

Inductie en Micro-CT beeldvorming van cerebrale Cavernous misvormingen in Mouse Model

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

Cardiale magnetische resonantie voor de evaluatie van vermoede cardiale trombus: conventionele en opkomende technieken

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...